03B Virology and Microbiology.pdf - Universidad AutÃ³noma de Madrid

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Virology and Microbiology 

Table of Contents 

Viral modulation of the immune response 

Antonio Alcami Pertejo 

Molecular bases of viral pathogenesis and anti-cancer potential 

José M. Almendral del Río 

Molecular Ecology of Extreme Environments 

Ricardo Amils Pibernat 

Bacterial Cell Division and Antibiotics Resistance 

Juan Alfonso Ayala Serrano 

Biotechnology and Genetics of Extreme thermophilic Bacteria 

José Berenguer Carlos 

ASFV: virus models of evasion and protection 

Ángel L. López Carrascosa 

Bacterial Morphogenesis 

Miguel Ángel de Pedro Montalbán 

Generic variability of RNA viruses 

Esteban Domingo Solans 

Gene expresión in Streptomyces and yeasts 

Antonio Jiménez / María Fernández Lobato 

Virus Engineering 

Mauricio García Mateu 

Effects of extrachromosomal elements on behaviour of its host Bacillus 

Wilfried J.J. Meijer 

Human immunodeficiency virus reverse transcriptase and antiretroviral therapy 

Luís Menéndez Arias 

African swine fever virus 

Maria Luisa Salas Falgueras 

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New strategies for prevention and control of viral diseases: 

foot-and-mouth disease virus as a model 

Francisco Sobrino 

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Virology 

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Research Summary 

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Línea Investigación 

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We are investigating immune evasion mechanisms employed by large DNA 

viruses, poxviruses and herpesviruses. Specifically, we are characterizing a unique 

immunomodulatory strategy that consists of the secretion of viral proteins that bind 

cytokines and chemokines, important mediators of the inflammatory and immune 

response. We work on two virus systems: (1) Herpesviruses like herpes simplex virus 

and human cytomegalovirus, which are human pathogens of clinical relevance; and (2) 

Poxviruses such as vaccinia virus, the smallpox vaccine, and variola virus, the causative 

agent of smallpox and the only viral disease eradicated as a result of a global vaccination 

campaign. Secreted cytokine decoy receptors are likely to contribute to the pathogenesis 

of variola virus, which was one of the most virulent viruses known by mankind. The immune 

modulatory activity of the viral cytokine receptors and their contribution to pathology are 

being addressed in the mousepox model. Mousepox is a smallpox-like disease caused by 

ectromelia virus, a natural mouse pathogen, and a classical model of viral pathogenesis. 

Viruses have optimized during millions of years of evolution their ability to manipulate the 

host immune response. Viruses offer a unique opportunity to develop their immune evasion 

strategies as novel therapeutic approaches to block the damage caused by an uncontrolled 

inflammatory response. In collaboration with Biotech Companies, we are working towards 

the development of viral immunomodulatory proteins as potential medicaments to treat 

human allergic and autoimmune diseases caused by immunopathological reactions. 

Viruses are the most abundant and diverse biological entities on Earth. We are interested 

in the molecular characterization of complex viral communities using new massive 

sequencing methodologies (Roche-454 Life Sciences). We have described for the first 

time the viral community in an Antarctic lake and are expanding these studies to other 

lakes along the Antarctic Peninsula and in the Arctic. We are also interested in using 

the new sequencing technologies to identify viruses associated with human pathologies, 

such as multiple sclerosis. 

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Figure 1. Mimicry 

of cytokines and 

cytokine receptors 

by herpesviruses 

and poxviruses. 

Línea Investigación 

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Figure 2. Metagenomic 

studies of the viral 

community in Antarctic 

lakes. 

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Group Leader: 

Antonio Alcami Pertejo 

Postdoctoral Fellows: 

Ali Alejo Herberg 

Abel Viejo Borbolla 

Alberto López Bueno 

Daniel Rubio Muñoz 

Elena Merino Rodríguez 

Soledad Blanco Chapinal 

Imma Montanuy Sellart 

Juan Alonso Lobo 

Graduate Students: 

Marcos Palomo Otero 

Sergio Martín Pontejo 

Nadia Martínez Martín 

Carla Mavián 

Haleh Heidarieh 

Technical Assistance: 

Rocío Martín Hernández 

Carolina Sánchez Fernández 

Visiting Scientists: 

Joanne Devlin (University of 

Melbourne, Australia) 

Julia Fahel (Trinity College, 

Dublin, Ireland) 

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Andrés G., Leali, D., Mitola, S., Coltrini, D., Camozzi, M., Corsini, M., Belleri, M., Hirsch, E., Schwendener, R. A., Christofori, Alcami, 

A. and Presta, M. (2009) A pro-inflammatory signature mediates FGF2-induced angiogenesis. J. Cell. Mol. Med. 13, 2083-2108. 

Waibler, Z., Anzaghe, M., Frenz, T., Schwantes, A., Pöhlmann, C., Ludwig, H., Palomo-Otero, M., Alcami, A., Sutter, G. and Kalinke, 

U. (2009) Vaccinia virus-mediated inhibition of type I interferon responses is a multi-factorial process involving the soluble type I 

interferon receptor B18 and intracellular components. J. Virol. 83, 1563-1570. 

Carson, C., Antoniou, M., Ruiz-Argüello, M. B., Alcami, A., Christodoulou, V., Messaritakis, I., Blackwell, J. M. and Courtenay, O. 

(2009) A prime/boost DNA/Modified vaccinia virus Ankara vaccine expressing recombinant Leishmania DNA encoding TRYP is safe 

and immunogenic in outbred dogs, the reservoir of zoonotic visceral leishmaniasis. Vaccine 27, 1080-1086. 

Alcami, A. and Saraiva, M. (2009) Chemokine binding proteins encoded by pathogens. Adv. Exp. Med. Biol. 666, 167-179. 

Poole, E., Groves, I., Ho, Y., Benedict, C., Alcami, A. and Sinclair, J. (2009) Identification of TRIM23 as a co-factor involved in the 

regulation of NFkB by the human cytomegalovirus gene product UL144. J. Virol. 83, 3581-3590. 

Alejo, A., Ruiz-Argüello, M. B., Viejo, A., Saraiva, M., Fernández de Marco, M. M., Salguero, J. and Alcami, A. (2009) Adaptation of 

a transient dominant selection method to the generation of ectromelia virus recombinants: in vivo analysis of ectromelia virus CD30 

deletion mutants. PLoS ONE 4(4):e5175. 

Alcami, A. and Viejo-Borbolla, A (2009) Identification and characterization of virus-encoded chemokine binding proteins. Methods 

Enzymol. 460, 173-191. 

Shang, L., Thirunarayanan, N., Viejo-Borbolla, A., Martin, A. P., Bogunovic, M., Marchesi, F., Unkeless, J. C., Ho, Y., Furtado, G. C., 

Alcami, A., Merad, M., Mayer, L. and Lira, S. A. (2009) Expression of the chemokine binding protein M3 promotes marked changes in 

the accumulation of specific leukocytes subsets within the intestine. Gastroenterology 137, 1006-1018. 

López-Bueno, A., Tamames, J., Velázquez, D., Moya, A., Quesada, A. and Alcami, A. (2009) High diversity of the viral community from 

an Antarctic lake. Science 326, 858-861. 

Alejo, A. and Alcami, A. (2010) Chemokine binding proteins as therapeutics. In: Leurs, R., Smit, M. and Lira, S. (eds) Chemokine 

Receptors as Drug Targets. Wiley-VCH, Weinheim, Germany, pp.359-374. 

Rubio, N., Palomo, M. and Alcami, A. (2010) Interferon a/b genes are upregulated in murine brain astrocytes after Theiler’s murine 

encephalomyelitis virus infection. J. Interf. Cyt. Res. 30, 253-262. 

Viejo-Borbolla, A., Muñoz, A., Tabares, E. and Alcami, A. (2010) Glycoprotein G from pseudorabies virus binds to chemokines with 

high affinity and inhibits their function. J. Gen. Virol. 91, 23-31. 

Fernandez de Marco, M. M., Alejo, A., Hudson, P., Damon, I. K. and Alcami, A. (2010) The highly virulent variola and monkeypox 

viruses express secreted inhibitors of type I interferon. FASEB J. 24, 1479-1488. 

Devlin, J. M., Viejo-Borbolla, A., Browning, G. F., Noormohammadi, A. H., Gilkerson, J. R., Alcami, A. and Hartley, C. A. (2010) 

Evaluation of immunological responses to a glycoprotein G deficient candidate vaccine strain of infectious laryngotracheitis virus. 

Vaccine 28, 1325-1332. 

Alcami, A. and Lira, S. A. (2010) Modulation of chemokine activity by viruses. Curr. Opin. Immunol. 22, 482-487. 

Alcami A. (2010) The interaction of viruses with host immune defenses. Curr. Opin. Microbiol. 13, 501-502. 

Viejo-Borbolla, A., Martin, A. P., Muniz, L. R., Shang, L., Marchesi, F., Thirunarayanan, N., Harpaz, N., Garcia, R. A., Apostolaki, M., 

Furtado, G. C., Mayer, L., Kollias, G., Alcami, A. and Lira, S. A. (2010) Attenuation of TNF-driven murine ileitis by intestinal expression 

of the viral immunomodulator CrmD. Mucosal Immunol. 3, 633-644. 

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Member of the Editorial Board of Virology 

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Member of the Editorial Board of Journal of Virology 

Advisor to the World Health Organization Advisory Committee on Variola Virus Research 


Editor of a special issue of Curr. Opi. Microbiol. on Host-microbe interacions: viruses, vol. 

13, August 2010. 

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Doctoral Theses 

The research of the laboratory is mainly focused in the molecular mechanisms operating 

in viral diseases and in the design of viruses as potential anti-cancer biological tools. 

We use as experimental models the parvovirus Minute Virus of Mice (MVM) and the 

alphavirus Sindbis virus (SV). Among those topics recently addressed, or under current 

research interest, the following two are within the main ones: (i) The regulation of MVM 

capsid assembly in human cancer cells. We have found that VP2, the major MVM capsid 

protein, is specifically phosphorylated by Raf-1 (Figure 1), a kinase playing key roles in 

the development of several common human cancers. The phosphorylation by a mutationactivated 

Raf-1 facilitates the nuclear transport of VP2 trimers (Figure 1B), a process 

essential for the viral maturation. This MVM assembly regulation (Figure 1C) may explain 

most aspects of the parvovirus natural tropism toward human cancer cells, and it does 

identify a relevant target for possible oncolytic virotherapies. (ii) Translation control of viral 

mRNA. Our working hypothesis is that translation of viral mRNA may be directing key 

aspects of virus cycle such as host range and evolution, tropism for cells and tissues, and 

pathogenesis. Our main goal is to understand how the PKR cellular kinase modulates 

replication of SV (Figure 2) and MVM through the phosphorylation of translation initiation 

factor 2 (eIF2). By means of systems biology, we are also characterizing the molecular 

mechanism used by SV to overcome the antiviral effect of PKR, which involves specific 

structures in viral mRNA and the use of alternative initiation factors supporting viral 

translation. 

The group of Begoña Aguado (*) is focused on Alternative Splicing as a factor contributing 

to complexity and diversity among primates and other mammals, through the generation 

of different transcripts, including chimeric and non-coding ones. We are also investigating 

alternative splicing alterations which can cause disease, such as Rheumatoid Arthritis and 

Myotonic Dystrophy. We are using new technologies such as nano Q-PCRq, microarrays 

and Massive Sequencing. 

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Figure 1. The Raf- 

1 kinase regulates 

MVM assembly. (A) 

T w o - d i m e n s i o n a l 

phosphopeptide map of 

MVM capsid subunits 

in vitro phosphorylated 

by Raf-1. (B) Trimers of 

the MVM capsid protein 

isolated from human 

and insect cells differ in 

their nuclear transport 

competence. (C) The role 

of Raf-1 in the nuclear 

translocation of MVM 

assembly intermediates. 

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Figure 2. The shut-off 

phenomenon in mouse brain 

tissues infected with Sindbis 

virus. Translation of cellular, 

but not of viral mRNAs, 

was inhibited due to eIF2 

phosphorylation. 

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José M. Almendral del Río 

Scientific Staff: 

Begoña Aguado Orea (*) 

Antonio Talavera Díez 

Iván Ventoso Bande 


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Elena Domingo Gil 

Esther Grueso Hierro 

Francisco Hernández-Torres (*) 

Graduate Student: 

Noelia Blanco Menéndez 

Maria Elizalde Arbilla 

René Toribio López 

Olatz Villate Bejarano (*) 

Alberto Rastrojo Lastras (*) 

Raquel López-Díez (*) 

Undergraduate Students: 

Marian Fernandez Estevez 

Julia Wienke (*) 

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Sanz MA, Castelló A, Ventoso I, Berlanga JJ, Carrasco L. (2009). Dual mechanism for the translation of 

subgenomic mRNA from Sindbis virus in infected and uninfected cells. PLoS One. 4(3):4772. 

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Blanco, AM, L Rausell, B Aguado, M Perez-Alonso, R Artero. (2009). A FRET-based assay for characterization 

of alternative splicing events using peptide nucleic acid fluorescence in situ hybridization. Nucleic Acids 

Research 37 e116. 

Riolobos, L., Valle, N., Hernando, E., Maroto, B., Kann, M., and J. M. Almendral. (2010). Viral oncolysis that 

targets Raf-1 signaling control of nuclear transport. J. Virol., 84, 2090-2099. 


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Ventoso, I., Berlanga, J.J., and J. M. Almendral. (2010). Translational control by the Protein Kinase R 

restricts Minute Virus of Mice infection: role in parvovirus oncolysis. J. Virol., 84, 5043-5051. 

Toribio R, Ventoso I. Inhibition of host translation by virus infection in vivo. (2010). Proc Natl Acad Sci 

USA. 107(21):9837-42. 

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Olatz Villate. Splicing en el MHC de clase III: caracterización y expresión de las isoformas 

del gen NFkBIL1. Estudio de su relación con artritis reumatoide. Directora, 

Begoña Aguado. 

René Toribio López. Cambios traduccionales que regula la interacción virus hospedador. 

Implicación en el desarrollo de virus oncolíticos. Director, Iván Ventoso 


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Two main lines of research are currently in progress in our group: 

Molecular Ecology of Extreme Environments: This area of research has the following 

objectives: 

- Acidophiles: conventional microbial ecology (enrichment cultures, isolation, physiology), 

molecular ecology (DGGE, FISH, CARD-FISH, cloning), molecular biology (genomics, 

metagenomics, proteomics, differential gene expression using DNA arrays) and 

biotechnology (control of bioleaching, specific metal sequestering and phytoremediation) 

of extreme acidic environments (Río Tinto basin, different acidic lakes of the Iberian Pyritic 

Belt, río Agrio (Argentina), volcanic areas of Island, Antartica), 

- Geomicrobiological characterization of extreme environments as habitability models of 

astrobiological interest: Tinto basin (Mars geochemical analogue), sulfide deposits from 

Antartica (Mars analogue), Tirez hypersaline lagoon (Europa analogue, in collaboration 

with I. Marín, associate professor of the Department of Molecular Biology), Uyuni salt lake 

(Europa analogue, in collaboration with I. Marín), volcanic areas of Island, permafrost 

areas of Alaska (Mars analogue). 

- Geomicrobiology of the Iberian Pyritic Belt (IPB) subsurface: different techniques are 

being implemented in the characterization of the subsurface bioreactor responsible of the 

extreme acidic conditions of Río Tinto. This work is done in collaboration with the Centro 

de Astrobiología (Origin Project IPBSL) 

- The line of microbial ecology of anaerobic environments directed by professor J.L. Sanz 

(UAM) is being developed in the new facilities that the Department of Molecular Biology 

has in the Biology Building. This collaborative work is centred in the anaerobic activities 

detected in the different model systems studied by our group (Tinto basin, IPB, Tirez 

lagoon). 

Micology, This area of research directed by Dr. Aldo González has the following 

objectives: 

- Molecular genetics and microbiology of Basidiomicetes (Pleurotus ostreatus as model 

system). 

- Use as filamentous fungi as a source of secondary metabolites, lignolytic enzymes and 

specific sequestering of toxic metals. 

- Control and elimination of fungi from air-indoor. 

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Figura 1. Specific Cr(III) sequestering acidophilic fungi. 

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Figura 2. Acidophilic photosynthetic biofilm. 

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Ricardo Amils Pibernat 


Aldo González Becerra 

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Posdoctorales: 

Moustafa Malki 

Monika Oggerin de Orube 

Lourdes Rufo Nieto 

Javier Ruiz Pérez 


Patxi San Martín Uriz 

Enrique Marín Palma 

Carlotta Vizioli 

Irene Sánchez Andrea 


Nuria Rodríguez González 

Catalina del Moral Juarez 


Linda Amaral (MBL, Woods Hole, USA) 

Jim Field (Arizona State University, USA) 

Alberto González Fiaren (NASA-Ames, USA) 

Eric Zettler (MBL, Woods Hole, USA) 

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Fairen, A.G., Dávila, A.F., Duport, L.G., Amils, R., McKay, C.P. (2009) Nature, 459: 398-400. 

Fairen, A.G., Schulze-Makuch, D., Rodríguez, A.P., Fink, W., Dávila, A., Uceda, E.R., Furfaro, R., Amils, R., 

McKay, C.P. (2009) Planet. Space Sci., 57: 276-287. 

Díaz-Garretas, B., Asensi, A., Rufo, L., Rodríguez, N., Sánchez-Mata, D., Amils, R., de la Fuente, V. (2009) 

Northeastearn Naturalist, 16 : 56-64. 

Carnicero, D., Díaz, E., Escolano, O., Rubinos, D., Ballesteros, O., Barral, M.T., Amils, R., García Frutos, 

F.J. (2009) Adv. Materials Research, 71-73: 677-680. 

Eugenio, M.E., Carbajo, J.M., Martín, J.A., González, and A.E. Villar, J.C. (2009) J. Basic Microbiol. 

49(5): 433-440. 

González-Toril, E., Aguilera, A., Souza-Egipsy, V., Diez, M., López-Pamo, E., Sánchez-España, J., Amils, 

R. (2009) Adv. Materials Research, 71-73: 113-116. 

Amils, R., González-Toril, E., Aguilera, A., Rodríguez, N., Fernández-Remolar, D., Díaz, E., García- 

Moyano, A., Sanz, J.L. (2009) Adv. Materials Research, 71-73: 13-19. 

García-Moyano, A., González-Toril, E., Amils, R. (2009) Adv. Materials Research, 71-73: 109-112. 

González-Toril, E; Amils, R; Delmas, RJ, et al. 2009. Biogeosciences, 6(1): 33-44. 

de la Fuente, V., Rufo, L., Rodríguez, N., Amils, R., Zuluaga, J. (2009) Biol Trace Elem Res, DOI 

10.1007/s12011-009-8471-1. 

Aguilera, A., Souza-Egipsy, V., González-Toril, E., Rendueles, O., Amils, R. (2010). Internat. Microbiol., 13: 

29-40. DOI: 10.2436/20,1501.01.109. 

González-Toril, E., Aguilera, A., Rodríguez, N., Fernández-Remolar, D., Gómez, F., Díaz, E., García- 

Moyano, A., Sanz, J.L., Amils, R. (2010) Hydrometallurgy, 104: 329-333. 

Cid, C., Garcia-Descalzo, L., Casado-Lafuente, V., Amils, R., Aguilera, A. (2010) Proteomics, 10: 2026- 

2036 DOI 10.1002/pmic.200900592. 

Souza-Egipsy, V., Aguilera, A., Mateo-Martí, E, Martín-Gago, J.A., Amils, R. (2010). Geomicrobiol. J., 27: 

692-706. 

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Lalueza, J., Rius, A., Puig, R., Martí, E., Martí, J.F., Rodríguez, N., Amils, R. (2010). Journal of American 

Leather Chemists Association, 105: 210-221. 

Amaral-Zettler, L., Zettler, E.R., Theroux, S.M., Palacios, C., Aguilera, A., Amils, R. (20010) ISME J. 

doi:10.1038/ismej.2010.101. 

Arana-Cuenca, A., Téllez Jurado A., Yagüe, S., Fermiñan, E., Carbajo, J.M., González, T., Domínguez, A., 

Villar, J.C., and González, A.(2010) Forest Systems 19 (2): 234-240. 

Eugenio, M.E., Santos, S.M., Carbajo, J.M., Martín, J.A., Martín-Sampedro, R., González, A.E., and Villar, 

J.C. (2010) Bioresources Technology 101(6): 1866-1870. 

Fairén, A.G., Chevier, V., Abramov, O., Marzo, G.A., Gavin, P., Davila, A.F., Tornabene, L.L. , Bishop, J.L., 

Roush, T.L., Gross, C., Kneissl, T., Uceda, E.R., Dohm, J.M., Schulze-Makuch, D., Rodríguez, J.A.P., Amils, 

R., McKay, C.P. (2010) PNAS (USA), doi/10.1073/pnas.1002889107. 

Fairen, A. G., Dávila, A.F., Lim, D., Bramall, N., Bonaccorsi, R., Zavaleta, J., Uceda, E.R., Stoker, C.,, 

Wierzchos, J., Amils, R., Dohm, J.M., Andersen, D., McKay, C. (2010) Astrobiology, 821- 843. DOI:10.1089/ 

ast. 2009.0440. 

Fernández-Remolar, D., Sánchez-Román, M., Amils, R. (2010) Sustainability, 2: 2541-2554, 

doi:10.3390/su2082541. 

Gómez; F., Mateo-Martí, E., Prieto.Ballesteros, O., Martín-Gago, J., Amils, R. (2010) Icarus, doi:10.1016/j. 

icarus.2010.05.027. 

Carbajosa, S., Malki, M., Caillard, R., López, M.F., Palomares, F.J., Martín-Gago, J.A., Rodríguez, N., 

Amils, R., Fernández, V.M., De Lacey, A.L. (2010) Biosensors and Bioelectronics, 26: 877-880. 

Fernández-Remolar, D., Prieto-Ballesteros, O., Gómez-Ortiz, D., Fernández-Sampedro, M., Sarrazin, P., 

Gailhanou, M., Amils, R. (2010) Icarus, 211: 114-138. 

Menor-Salván, C., Tornos, F., Fernández-Remolar, D., Amils, R. (2010) Earth Planet. Sci. Lett., doi: 

10.1016/j.epsl.2010.09.020. 

Capítulos de libros: Cinco capítulos de libro. 

Libros: Amils, R., Segura, J. (2010) Río Tinto… viaje a Marte, ediciones Alfar (Sevilla). 

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Amils, R. Malki, M., Fernández, V., de Lacey, A.L. 

Título: Electrodo bacteriano aeróbico para ánodo de una pila de combustible sin mediadores 

redox ni membrana intercambiadora de protones. Nº de solicitud: 200701534. Fecha de 

concesión: 23/12/2009 


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Eric Zettler, enero 2009, The relationship between environment and the microbial 

community in a patchwork of geochemical “islands”, Ricardo Amils, Universidad Autónoma 

de Madrid. 


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The objectives of our group are the analysis of the bacterial growth and cell division under 

diverse molecular approaches, and to study the mechanisms of resistance to the β-lactam 

antibiotics, that have been developed by pathogens of clinical origin. Under this general 

scheme, we have developed two main aspects. One is the molecular characterization of 

the CTX-M β-lactamases family, their mobilization mechanisms and the ancestral origin 

of the CTX-M-1 subfamily. Also the identification of PBPs and their role on the resistance 

mechanisms of anaerobic strains and Gram-negative enteric bacteria was analyzed. A new 

family of PBPs, belonging to the COG1680, with high homology with class C β-lactamases 

and been involved in morphogenesis, are going to be analyzed in a new project. 

Based on analysis of peptidoglycan structure of Aeromonas PBP4 mutants, we have 

proposed a model for induction of the expression of β-lactamase mediated by a two 

component regulatory system. (See joint figure) 

During cell division, assembly of proteins at a division ring has the effect of constricting 

the membrane and producing a cell wall septum. The synthesis of a rigid peptidoglycan 

septum involves a set of dedicated enzymes, as penicillin-binding proteins. We have 

achieved the purification of several of these enzymes (PBP1B, PBP3, and inactive 

variants) together with a good number of substrates (including sacculi, as the largest 

available structure, fragmented peptidoglycan and smaller-sized precursors as lipid II and 

UDP-muramyl pentapeptide) to assist in the set up of “in vitro” screening assays. 

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Figure 1. Model for beta-lactamase induction in 

Aeromonas spp. and the role of the blr regulon. 

Inhibition of PBPs by b-lactams causes an increase 

in the concentration of the BlrB activatory ligand. This 

ligand then interacts with BlrB (dotted arrow), causing 

it to auto-phosphorylate (‘P’ in the diagram). This 

phosphate is transferred to BlrA, which binds to the cre/ 

blr tag sequence found upstream of all blr regulon gene 

promoters. The effect of this is to recruit RNA polymerase 

and activate blr regulon transcription. Known blr regulon 

genes encode three blactamases, Amp, Cep and Imi, 

which directly hydrolyse the b-lactam, helping to prevent 

further peptidoglycan damage. The blr regulon is also 

known to include non-b-lactamase-encoding genes, e.g. 

blrD, and it is proposed that their products have a role in 

protecting the cell from b-lactam challenge through an 

ancillary mechanism. 

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Figure 2. DD-endopeptidase activity of AmpH identified 

by HPLC. Peaks correspond to M4.- NAcGlc-NAcMurtetrapeptide 

, M5.- NAcGlc-NAcMur-pentapeptide, D45.- 

Disacharide-tetra-pentapeptide, and C.- cefmetazol. 

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Juan Alfonso Ayala Serrano 

Postdoctoral Fellow: 

Silvia Marina González Leiza 

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Cristian Gustavo Aguilera Rossi 

Alaa Ropy Mahmoud Sayed 


Godofrey Cherry 

Yasemin Ezgi Ertürk 

Andrés Caballero 

Laura Vuolo 

Jose Roberto Angeles Vázquez 

Nicolas Cordeiro 

Ana Fernández González 

Sorelis Urdaneta Fernández 

Lucia Lozano 

Paula Andrea Espinal 

María Margarita Bernal 


Ayelen Patricia Porto (Universidad de Buenos 

Aires, Buenos Aires, Argentina) 

Ana Catarina Souza Lopes (Universidade 

Federal de Pernambuco, Brasil) 

Bartolome Moya (Universidad Islas Baleares, 

Mallorca) 

Judith J. Velasco (Universidad de los Andes, 

Mérida, Venezuela) 

-József Sóki University of Szeged, 

-Elizabeth Nagy University of Szeged, 

-Gabriella Terhes 

(University of Szeged, Szeged, Hungary) 

Sergei Borchsenius (Institute of Cytology, 

RAS, Saint-Petersburg, Russia) 

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Publications 

D. Korsak, Z. Markiewicz, G. O. Gutkind, and J. A. Ayala. (2010) Identification of the full set of Listeria 

monocytogenes penicillin-binding proteins and characterization of PBPD2 (Lmo2812). BMC Microbiology, 

10:239-251 

Inés Bado, Nicolás F. Cordeiro, Luciana Robino, Virginia García-Fulgueiras, Verónica Seija, Cristina Bazet, 

Gabriel Gutkind, Juan A. Ayala, and Rafael Vignoli. (2010). Detection of class 1 and 2 integrons, extendedspectrum 

β-lactamases and qnr alleles in enterobacterial isolates from the digestive tract of Intensive Care 

Unit inpatients. Intern. J. Antimicrob. Agents, 36(5):453-458 

A.C.S. Lopes, D. Leal Veras, A.M.S. Lima, R.C. Andrade Melo, and J.A. Ayala. (2010). bla(CTX-M-2) and 

bla(CTX-M-28) extended-spectrum beta-lactamase genes and class 1 integrons in clinical isolates of 

Klebsiella pneumoniae from Brazil.. MIOC. 105(2):163-167. 

Amy E. Tayler, Juan A. Ayala, Pannika Niumsup, Katrin Westphal, Timothy R. Walsh, Bernd Wiedemann, 

Peter M. Bennett, and Matthew B. Avison. (2010). Induction of beta-lactamase production in Aeromonas 

hydrophila is responsive to beta-lactam-mediated changes in peptidoglycan composition. Microbiology. 

156(Pt 8):2327-35. 

Porto, Ayelén; Ayala, Juan; Gutkind, Gabriel; and Di Conza, José. (2010). A novel OXA-10-like β-lactamase 

is present in different Enterobacteriaceae. Diagnostic Microbiology and Infectious Disease. 66:228-229. 

M. Macedo-Viñas, N. F. Cordeiro, I. Bado, S. Herrera-Leon, M. Vola, L. Robino, R. Gonzalez-Sanz, S. 

Mateos, F. Schelotto, G. Algorta, J. A. Ayala, A. Echeita and R. Vignoli. (2009). Surveillance of antibiotic 

resistance evolution and detection of class 1 and 2 integrons in human isolates of multiple resistant 

Salmonella Typhimurium obtained in Uruguay from 1976 to 2000. International Journal of Infectious 

Diseases, 13:342-348. 

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Profesor “Ad Honorem”, Facultad de Medicina, Universidad de la Republica, Uruguay 

Julio, 2009. 


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We use the extreme thermophilic bacteria Thermus thermophilus (Tth) as our main 

laboratory model. Tth grows quite fast at 70 ºC, forms colonies on plates, and shows natural 

competence. As other thermophiles, its enzymes show a greater ability to crystallize than 

their mesophilic homologues, thus making it an excellent model for Structural Biology. Its 

ancient phylogenetic origin also adds biological and evolutionary interest to this model. 

We pursue both biotechnological and basic-science objectives with this model. In 

biotechnology: i) We overproduce and use thermostable enzymes (thermozymes) for 

biotransformations in collaboration with external groups specialized in biocatalysis; ii) 

We evolve enzymes and proteins towards either increasing their thermostability through 

in vivo selection, or their specific activities at low temperatures; and iii) We develop new 

genetic tools for its use in thermophiles. In the last two years we have overproduced 

more than 50 thermozymes (dehydrogenases, estherases, and glycosidases) and carried 

out selection procedures for the isolation of thermostable mutants of three proteins. 

In basic-science we study the energy metabolism of Tth, specifically the respiration of 

nitrogen oxides –denitrification-, its genetic control, and its lateral gene transfer (LGT). 

Through massive sequencing we have shown that the process is catalyzed by three 

reductases encoded within a denitrification island (DI) that is the subject of frequent LGT 

events. This DI concentrates information on new type of enzymes that replace the aerobic 

respiratory complexes I and III, by more simple ones or by bifunctional enzymes acting as 

reductases-electrons transporter. The DI also encodes new sensory-signal transduction 

system that allows the organism to select the components of its respiratory chains 

according to environmental signals. For the next years we will continue these studies 

and we start new projects on unconventional mechanisms that prevent the LGT between 

bacterial species. 

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Figure 1. The denitrification 

apparatus of Thermus 

thermophilus. An scheme of 

the localization and role of the 

NADH dehydrogenase (Nrc), 

and the nitrate (Nar), nitrite 

(Nir), and nitric oxide (Nor) 

reductases is shown. Note 

the role of Nar as electron 

transporter replacing the 

complex III. Electrons pathway 

is shown as red arrows. 

Staff 

Publications 

Patents 

Figure 2. Evolving an 

esterase. (R)-1-phenyl- 

2-propyl acetate docked 

in the active site of a 

quadruple mutant (mutant 

Q) of the Pseudomonas 

fluorescens esterase I. 

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José Berenguer Carlos 


Aurelio Hidalgo Huertas 


Alba Mº Sanchez Niño, 

Akbar Espaillat Fernández 

Ángel Cantero Camacho 

Daan Swarts 


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Publications 

Patents 


Daniel Vega 

Leticia Torres 


Federico Acosta 

Eloy Ferreras Puente 

Zahra Chahlafi 

Marcos Almendros Gimenez 

Laura Álvarez Muñoz 

Carlos Bricio Garberí 

Noé Rigoberto Rivera 

Yamal Al-Ramahi González 

Martin Hesseler 


Esther Sánchez Freire 

Maria José de Soto López 


Estariette van Heerden 

Koos Albertin 

Derek Litthauer 

Godfrey Tlou 

Philip Armand Bester 

Mariana Erasmus 

Novalanda Betty Mabizela 

Susana Alarico 

Hernán Costa 

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Publications 

Larsen,M., Zielinska, D.F., Martinelle, M., Hidalgo, A., Jensen, L., Bornscheuer, U.T. and Hult, K. (2010) 

Suppression of water as a nucleophile in Candida antarctica lipase B catalysis. ChemBioChem 11,796-801. 

Cava, F, Hidalgo, A., and Berenguer, J. (2009) Thermus thermophilus as biological model. Extremophiles, 

13, 213-231. 

Schließmann, A., Hidalgo, A., Berenguer, J. and Bornscheuer, U.T (2009). Increased Enantioselectivity by 

Engineering Bottleneck Mutants in an Esterase from Pseudomonas fluorescens. ChemBioChem 10,2920- 

2923 (cover feature). 

Rocha-Martin, J., Vega D., Cabrera Z, Bolivar JM. Fernandez-Lafuente R., Berenguer J. and Guisan, J.M. 

(2009) Purification, immobilization and stabilization of a highly enantioselective alcohol dehydrogenase 

from Thermus thermophilus HB27 cloned in E. coli. Proccess in Biochemistry, 44,1004-1012. 

Bolivar J.M., Rocha-Martin J., Mateo C., Cava F., Berenguer, J, Vega D. , Fernandez-Lafuente R., Guisan 

J.M. (2009) Purification and stabilization of a glutamate dehygrogenase from Thermus thermophilus via 

oriented multisubunit plus multipoint covalent immobilization. Journal of Molecular Catalysis B: Enzymatic 

58, 158-163. 

Almendros M., Sinisterra JV, and Berenguer J. (2009) Thermus thermophilus Strains Active in Purine 

Nucleoside Synthesis. Molecules 2009, 14, 1279-1287. 

Bolivar J.M, Rocha-Martin J. Mateo C., Cava F., Berenguer J., Fernandez-Lafuente R., Guisan J. M. 

(2009) Coating of soluble and immobilized enzymes with ionic polymers: full stabilization of the quaternary 

structure of multimeric enzymes. Biomacromolecules 10, 742-747 

Bolivar J., Mateo, C., Rocha-Martin, Cava F., Berenguer J., Fernandez-Lafuente R., Guisan J. M. 2009. 

The adsortion of multimeric enzymes on very lowly activated supports involves more enzyme subunits: 

stabilization of glutamate dehydrogenase from Thermus thermophilus by immobilization on heterofunctional 

supports. Enzyme Microb. Technol. 44, 139-144. 

Cava, F., Chahlafi, Z., Alavare, L., and Berenguer, J. (2009) Respiración y desnitrificación en Thermus 

thermophilus. In: Bonete, M. J., and Martínez-Espinosa R. M. (ed) Avances en el metabolismo del nitrógeno. 

Editorial Club Universitario, Alicante. pp. 173-186. 

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Procedimiento de inmovilización de una glutamato deshidrogenasa. 

PCT/ES2008/070166. 

Nuevo marcador termoestable para la selección genética de Thermus spp P200603279. 

(Concesión 28-05-2010). 


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During the last years we have been searching for established cell lines sensitive to the 

African Swine Fever Virus (ASFV), for the production and titration of virus isolates both 

from the field or laboratory, as well as for the generation of virus recombinants with specific 

genes deleted. These deletion mutants facilitate the study of the role of several ASFV 

genes involved in the evasion of the antiviral response of the cell during virus infection, 

and may serve as attenuated virus models in the generation of possible vaccines to induce 

protective immunity against ASF. So, we have confirmed the role of the ASFV lectin 

(EP153R gene) in the modulation of the expression of MHC-I antigens in the membrane 

of the infected cell, and we are also inactivating several viral genes in a partially-attenuated 

ASFV isolate (NHV), able to induce protection against the virulent L60 isolate, but still 

retaining a residual virulence unacceptably high to be used as a vaccine. 

The porcine cell lines sensitive to ASFV infection are also being used for the determination 

of the level of induction of selected cytokines by ASFV isolates with various degrees 

of virulence, searching for a possible “virulence/attenuation profile”, which might result 

in a remarkable reduction of the in vivo infections required to determine the degree of 

virulence of the virus isolates. 

Another objective of our Group is the optimization of the use of non-conventional antivirals 

as the lauryl gallate (LG) in the prevention and/or treatment of viral diseases of clinical 

and veterinarian importance. We have demonstrated its low cytotoxicity and its efficiency 

in the inhibition of the virus production in several models (ASFV, influenza, herpes,...) in 

cells infected in the laboratory, and we pretend to extend the study of the protective level 

provided by the LG in in vivo infections. 

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Figure 1. Interaction EP153R - SLA-I. Residues involved 

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Ángel L. López Carrascosa 

Postdoctoral: 

Patricia de León Valdés 


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Undergraduated Student: 

Alba Martínez Flórez 


Maria José Bustos Sánchez 

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Hurtado, C., Bustos, M. J. and Carrascosa, A. L. (2010). The use of COS-1 cells for studies of field and laboratory 

African swine fever virus samples. J. Virol. Methods 164, 131-134. 

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Hurtado, C., Bustos, M. J., Granja, A. G., de León, P., Sabina, P., Lopez-Viñas, E., Gomez-Puertas, P., Revilla, 

Y., and Carrascosa, A. L. (2010). The African swine fever virus lectin EP153R modulates the surface 

membrane expression of MHC class I antigens. Arch. Virol. DOI: 10.1007/s00705-010-0846-2. 


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Bacterial morphogenesis 

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Our main interest is the investigation, at the molecular and structural levels, of the bacterial 

cell wall (sacculus or peptidoglycan layer) as the primary morphogenetic element of 

the prokaryotic cell. We are presently working on cell development, adaptation and 

differentiation of appendages (prostheca, flagella, pili and hold-fast) in alfa-proteobacteria, 

mainly in Asticcacaulis biprosthecium, and other bacterial groups of complex morphology. 

We want to extend our experience in simple cell-cycle organisms as Escherichia coli, 

to more complex bacteria as A. biprosthecium. This species presents a dimorphic cell 

cycle with a mobile flagellated form (swarmer cell) which eventually differentiates into a 

sessile form displaying two laterally opposed prostheca. Only the sessile form is division 

proficient, and generates a sessile and a swarmer cell upon division. At present we are 

investigating how the metabolism and structure of the cell wall are modified in accordance 

with the cell cycle differentiation processes, and in relation with the generation of cell 

appendages (prostheca, flagella, and hold-fast) at precise places and times in the cell 

cycle. Recently we started a new line of work centred on the discovery of a novel regulatory 

mechanism of cell wall metabolism in Vibrio cholerae and other bacteria. The mechanism 

is mediated by the production and release of specific D-amino acids, is part of the general 

stress response mechanism, and has the potential to act as an inter and intra specific 

signalling system. We are also involved in cooperative projects concerning structural and 

biochemical aspects of cell wall metabolism in other organisms, Listeria monocitogenes 

or Leptospira biflexa, associated to the development of symbiotic or pathologic relations 

and to peculiar morphologies. 

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Figure 1. Asticcacaulis biprosthecium cells 

visualized by confocal microscopy. 

Figure 2. Actively growing (A) and resting (B) 

cells of Vibrio cholera as visualized by ESM. 

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Miguel Angel de Pedro Montalbán 


Said Taimani 



Marisela Domínguez Domínguez 

María Hidalgo García 

Irene Cartas 

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Lam, H, Oh, D.C., Cava, F., Takacs, C.N., Clardy, J., de Pedro, M.A. and Waldor, (2009) D-amino acids 

govern stationary phase cell wall remodeling in bacteria. Science 325:1552-1555. 

de Pedro, M.A. (2009). Peptidoglycan (Murein), In Encyclopedia of Microbiology. (Moselio Schaechter, 

Ed.), pp. 453-469, Oxford: Elsevier. 


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B8 

Genetic variability of RNA viruses 

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The main interest of our group is to understand the molecular bases of RNA virus 

extinction by lethal mutagenesis, and to explore antiviral protocols using mutagenic 

agents and antiviral inhibitors. We have documented a double mutagenic and inhibitory 

activity of 5-fluorouracil on foot-and-mouth disease virus (FMDV), and have characterized 

FMDV mutants resistant to the mutagenic purine analogue ribavirin. Specifically, we 

have described an amino acid substitution in the FMDV polymerase which avoids the 

deleterious activity of ribavirin by means of a modulation of the mutation types produced 

by the drug. Virus survival through modulation of mutation types, without an alteration 

of the general copying fidelity of the polymerase, constitutes a new mechanism of 

resistance to a mutagenic agent. Despite the presence of mutagen-resistance mutations, 

virus extinction can be achieved with alternative mutagenic treatments. Contrary to what 

has been established for classic antiviral treatments, a sequential administration of an 

inhibitor first, and then of a mutagenic agent can be more effective for virus extinction 

than the administration of the two drugs simultaneously. 

With regard to the understanding of the biological implications of quasispecies behavior, we 

have characterized a competition-colonization dynamics among FMDV subpopulation that 

arose in cell culture through diversification of a biological clone of the virus. Our laboratory 

has participated in collaborations with various teams on theoretical and experimental 

virology, on structural studies with the FMDV polymerase, and biological implications of 

quasispecies, including new vaccine designs. Such collaborations can be identified by the 

names of the authors of the publications included in the present summary. 

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Figure 1. Antiviral strategy based on the extinction of viruses by increasing their mutation 

rates during replication. 

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Esteban Domingo Solans 


Celia Perales Viejo 

Verónica Martín García 

Armando Arias Esteban 

Julie Sheldon 


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Publications 


Rubén Agudo Torres 

Marta Sanz-Ramos Rojo 

Samuel Ojosnegros Martos 

Héctor Moreno Borrego 

Héctor Tejero Franco 

Ignacio de la Higuera 

Ana Mª Ortega Prieto 

Patents 



Ana Isabel de Ávila Lucas 

Eva García Cueto 

Isabel Gallego Jiménez 

Mª Eugenia Soria Benito 

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Agudo, R. Arias, A. and Domingo, E. (2009). 5-Fluorouracil in lethal mutagenesis of foot-and-mouth disease virus. Future Medicinal 

Chemistry, 1(3), 529-539. 

Domingo, E. and Wain-Hobson, S. (2009). The 30th anniversary of quasispecies. Meeting on quasispecies: past, present and future. 

EMBO Reports, 10, 444-448. 

Emonet, S.F., de la Torre, J.C., Domingo, E. and Sevilla, N. (2009). Phylogeny/molecular taxonomy and evolution or Arenaviruses. Infection Genetics 

and Evolution, 9, 417-429. 

Perales, C., Agudo, R. and Domingo, E. (2009). Counteracting quasispecies adaptability: extinction of a ribavirin resistant virus mutant by an 

alternative mutagenic treatment. PLoS One, 4(5): e5554.doi: 10.1371/journal.pone.0005554. 

Escarmís, C., Perales, C. and Domingo, E. (2009). Biological effect of Muller’s ratchet. Distant capsid site can affect picornavirus 

protein processing. J. Virol. 83, 6748-6756. 

Domingo, E. (2009). Nueva gripe humana de origen porcino. ¿Cuánto de nuevo? Investigación y Ciencia. 393, 10-12. 

Ferrer-Orta, C., Agudo, R., Domingo, E. and Verdaguer, N. (2009). Structural insights into replication and elongation processes by the FMDV 

RNA-dependent RNA polymerase. Curr. Op. in Structural Biol. 19 (6), 752-8. 

Perales, C., Agudo, R., Tejero, H., Manrubia, S.C. and Domingo, E. (2009). Potential benefits of sequential inhibitor-mutagen treatments of RNA virus 

infections. PLoS Pathogens. 5 (11), e1000658. 

Domingo, E. (2009). Quasispecies. From molecular Darwinism to viral diseases. Contributions to Science. 5(2), 161-168. 

Ojosnegros, S., Beerenwinkel, N., Antal, T., Nowak, M.A., Escarmís, C. and Domingo, E. (2010). Competition-colonization dynamics in an RNA virus. 

Proc. Natl. Acad. Sci. USA. 107(5), 2108-12. 

Domingo, E. (2010). The great evolutionary potential of viruses. The 1918 flu as a paradigm of disease emergence. In: M.I. Porras and R. Davies, 

ed. Emerging Infection, Emergent Meanings: The “Spanish” Influenza Pandemic of 1918-1919. University of Rochester Press, Rochester, New York. 

107, 2108-2112. 

Domingo, E. (2010). Mechanisms of viral emergence. Veterinary Research, 41(6): 38. 

Martín, V. Abia, D., Domingo, E. and Grande-Pérez, A. (2010). An interfering activity of LCMV associated with enhanced mutagenesis. J. 

Gen. Virol., 91, 990-1003. 

Ojosnegros, S., Beerenwinkel, N. and Domingo, E. (2010). Competition-colonization dynamics: an ecology approach to quasispecies dynamics and 

virulence evolution in RNA viruses. Communicative & Integrative Biology, 3 (4), 333-6. 

Domingo, E., Perales, C., Agudo, R., Arias, R., Escarmís, C., Ferrer-Orta, C. and Verdaguer, N. (2010). Mutation, quasispecies and lethal mutagenesis. 

In: E. Ehrenfeld, E. Domingo and R.P. Roos, eds. The Picornaviruses. ASM Press, Washington, D.C., pp.197-211. 

Manrubia, S.C, Domingo, E. and Lázaro, E. (2010). Pathways to extinction: beyond the error threshold. Phil. Trans. R. Soc. B., 365(1548), 1943-52. 

Ferrer-Orta, C., Sierra, M., Agudo, R., de la Higuera, I., Arias, A., Pérez-Luque, R., Escarmís, C., Domingo, E. and Verdaguer, N. (2010). Structure of 

foot-and-mouth disease virus mutant polymerases with reduced sensitivity to ribavirin. J. Virol., 84(12), 6188-99. 

Bordería, A.V., Lorenzo-Redondo, R., Pernas, M., Casado, C., Álvaro, T., Domingo, E. and López-Galíndez, C. (2010). Initial fitness recovery of HIV-1 

is associated with quasispecies heterogeneity and can occur without modifications in the consensus sequence. PLoS One, 5(4), e10319. 

Martín-Acebes, M.A., Herrera, M., Armas-Portela, R., Domingo, E. and Sobrino, F. (2010). Cell density-dependent expression of viral antigens during 

persistence of foot-and-mouth disease virus in cell culture. Virology, 403, 47-55. 

Rodríguez-Calvo, T., Ojosnegros, S, Sanz-Ramos, M., García-Arriaza, J., Escarmís, C., Domingo, E. and Sevilla, N. (2010). New vaccine design 

based on defective genomes that combines features of attenuated and inactivated vaccines. PLoS One, 5(4), e10414. 

Arias, A., Perales, C., Escarmís, C. and Domingo, E. (2010). Deletion mutants of VPg reveal new cytopathology determinants in a picornavirus. 

PLoS One, 5(5), e10735. 

Agudo, R., Ferrer-Orta, C., Arias, A., de la Higuera, I., Perales, C., Pérez-Luque, R., Verdaguer, N. and Domingo, E. (2010). A multi-step process of 

viral adaptation to a mutagenic nucleoside analogue by modulation of transition types leads to extinction-escape. PLoS Pathog. 6(8) pii: e1001072. 

Perales, C., Lorenzo-Redondo, R., López-Galíndez, C., Martínez, M. A., and Domingo, E. 2010. Mutant spectra in virus behavior. Future 

Virology 5(6), 679-698. 

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N. Sevilla, E. Domingo, C. Escarmís, S. Ojosnegros, J. García-Arriaza, M. Sanz-Rojo, T. 

Rodríguez (2009) Vacuna atenuada para la fiebre aftosa. PCT1641.49. Entidades titulares: 

CSIC (70%), Instituto Nacional de Investigaciones Agrarias (30%). 

E. Domingo, R. Agudo, H. Tejero, S.C. Manrubia, C. Perales (2009) Tratamiento antiviral. 

P200930482. Entidades titulares: CSIC (41%), Instituto Nacional de Técnica Aeroespacial 

(18%), Universidad Complutense de Madrid (18%), Centro de Investigaciones Biomédicas 

en Red de Enfermedades Hepáticas y Digestivas (CIBERehd) (23%). 


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Samuel Ojosnegros Martos (2009) Dinámica evolutiva de virus RNA. Universidad Autónoma 

de Madrid. Director: Esteban Domingo Solans. 

Rubén Agudo Torres (2009) Caracterización de las proteínas del virus de la fiebre aftosa 

implicadas en respuesta a mutagénesis letal por análogos de nucleótido. Universidad 

Autónoma de Madrid. Director: Esteban Domingo Solans. 


Marta Sanz-Ramos Rojo (2009) Dinámica de cuasiespecies del virus de la fiebre aftosa 

in vivo. Universidad Autónoma de Madrid. Director: Esteban Domingo Solans. 

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B9 

Gene expression in Streptomyces and yeasts 

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Streptomyces and yeasts produce a high number of molecules with biotechnological value. 

Our objective is to study the expression molecular mechanism of proteins implicated in 

some of this biosynthetic process, and to obtain new molecules with possible industrial 

applicability. During the last two years, we have been studying the biosynthetic cluster 

of the nucleoside antibiotic A201A (ata) from Streptomyces capreolus, and some yeast 

proteins with glycosyltransferase activity able to produce prebiotic oligosaccharides. 

The glycosyl residues constitute about 44% of the A201A total mass, which must be related 

with the biological-pharmacological properties of the antibiotic. We have characterized 

some genes involved in the incorporation and modification of these residues using different 

heterologous expression systems. The biosynthesis of novel molecules with antibiotic 

activity will be attempted based on the low substrate specificity previously described for 

the related glycosyltransferases. 

We are studying several proteins from the Xanthophyllomyces, Schwanniomyces and 

Rhodotorula yeasts genera, which produce different types of prebiotic oligosaccharides. 

All the analysed proteins are hydrolases that show transferase activity, included within 

family 32, 31, 1 and 2 of the glycosylhydrolases. To know the structure-function-specificity 

relationships of these proteins are one of our objectives. We are carrying out structural 

studies of these proteins (3-D structure of same of them has been resolved), residue 

substitutions, and applying directed molecular evolution techniques in order to increase/ 

modify their transglycosylase activity, and to improve their biotechnological utility. 

International patents of most of these proteins have been obtained and a method for its 

immobilization on solid support has been developed. We seek to scale up to industrial 

level the enzyme production and the generated products. 

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Figure 1. (A) Close up view of the 

active site in Schwanniomyces 

occidentalis Ffase in a complex 

with fructose, 1-kestose 

(green; left) and the 6-kestose 

(yellow; right). (B) The fructan 

1-exohydrolase IIa from Cichorium 

intybus (CiFEH) complexed with 

1-kestose and (C) the invertase 

from Thermotoga maritima 

complexed with raffinose. In (A), 

the active-site of Ffase (blue) 

is also shaped by the adjacent 

subunit (orange). 

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Figure 2. Oligomerization 

pattern of the Ftase 

from Schwanniomyces 

occidentalis on a this 

yeast culture. 

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Antonio Jiménez / María Fernández Lobato 

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Miguel de Abreu Felipe 

Miguel Álvaro Benito 

Dolores Linde López 

Patricia Gutiérrez Alonso 

Brian Molloy Galiana 


David González Pérez 

Marta Estévez Canales 

Hugo Muñoz 


Asunción Martín Redondo 


Víctor Cifuentes (Chile) 

Marcelo Baeza (Chile) 

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Publications 

Linde, D., Macias, I., Fernández-Arrojo, L., Plou, F.J., Jiménez, A. and Fernández-Lobato, M. (2009) 

Molecular and biochemical characterization of an fructofuranosidase from Xanthophyllomyces dendrorhous. 

App Env Microbiol. 75(4), 1065-1073. 

Gutiérrez-Alonso, P., Fernández-Arrojo, L., Plou, F.J. and Fernández-Lobato, M. (2009) Biochemical 

characterization of a b-fructofuranosidase from Rhodotorula gracilis with transfructosylating activity. FEMS 

Yeast Research. 9, 768-773. 

Polo, A., Álvaro-Benito, M., Fernández-Lobato, M. and Sanz-Aparicio, J. (2009) Crystallization and 

preliminary X-ray diffraction analysis of thefructofuranosidase from Schwanniomyces occidentalis. Acta 

Cryst. 65, 1162-1165. 

Baeza, M., Retamales, P., Sepulveda D., Lobato P., Jimenez A. and Cifuentes V. (2009) Isolation, 

characterization and long term preservation of mutant strains of Xanthophyllomyces dendrorhous. J. Basic 

Microbiol. 49, 135-141. 

Alvaro-Benito, M., Polo, A., González, B., Fernández-Lobato, M. and Sanz-Aparicio, J. (2010) Structural 

and kinetic analysis of Schwanniomyces occidentalis invertase reveals a new oligomerization pattern and 

the role of its supplementary domain in substrate binding. J. Biol. Chem. 285 (18), 13930-14941. 

Polo, A., Linde, D., Estévez, M., Fernández-Lobato, M. Julia Sanz-Aparicio, J.. (2010). Crystallization and 

preliminary X-ray diffraction analysis of the fructofuranosidase from Xanthophyllomyces dendrorhous. Acta 

Cryst. 66, 1441-1444. 

Álvaro-Benito, M., de Abreu, M., Portillo, F., Sánz-Aparicio, J. and Fernández-Lobato, M. (2010). New 

insights into the fructosyltransferase activity of Schwanniomyces occidentalis β-fructofuranosidase emerges 

from a non-conventional codon usage and directed mutation. App. Env. Microbiol. 76 (22), 7491-7499. 

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L. Fernández Arrojo, F. Plou, A. Ballesteros, M. Alcalde, P. Gutierrez Alonso, M. Fernández- 

Lobato (2009). Biocatalizador inmovilizado basado en alginato para la biotransformación de 

carbohidratos. Nº P200930001. PCT-ES2010/070104 (24 -2- 2010). Titular: CSIC-UAM. 

M. Fernández-Lobato, M. Alvaro Benito (2009). Fructofuranosidasa mejorada genéticamente 

para la obtención del prebiótico 6-kestosa. Nº P0200930929 (29-12-2009). Titular: UAM. 


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Brian Molloy Galiana (2010). Expresión heteróloga del cluster biosintético del antibiótico 

A201A y estudio de las posibles glicosiltransferasas implicadas en su biosíntesis. 

Universidad Autónoma de Madrid. Director: María Fernández-Lobato. 

María Dolores Linde López (2010). Caracterización bioquímica, molecular y estructural 

de una b-fructofuranosidasa con capacidad trasferasa de la levadura Phaffia rhodozyma 

aplicable a la producción de oligosacáridos prebióticos. Universidad Autónoma de Madrid. 

Director: María Fernández-Lobato. 

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We use protein engineering techniques for the study of structure-function relationships 

in viruses and their capsids, and for the design of modified viral particles for 

nanobiotechnological applications (Mateu (2011). Prot.Eng.Des.Sel. 24, 53-63). 

Scientific relevance and technological implications: A better knowledge of poorly 

known stages of the viral cycle, including virus assemby, conformational dynamics and 

disassembly; design of new vaccines, antivirals and nanoparticles for targeted drug 

delivery. 

Some recent scientific results: i) Using protein engineering, the thermal stability of footand-mouth 

disease virus (FMDV) against dissociation into subunits has been increased. 

We are currently investigating the molecular basis of such thermostabilization, and the 

possibility of using the modified virions as improved vaccines. ii) We have carried out an 

extensive mutational analysis of compensatory mutations in the FMDV capsid, and are 

currenly investigating the molecular bases of these effects. iii) In collaboration with P.J. de 

Pablo y J. Gómez (Dept. Physics of Condensed Matter, UAM) we have used the minute 

virus of mice (MVM) to investigate the role of capsid pores and cavities in the mechanical 

properties of the viral particle, using atomic force microscopy (a technique we are currently 

using in our lab). We have obtained evidence that the remarkable mechanical properties 

found may have been evolutionarily selected to optimize the resistance of the virus against 

physical agents, while at the same time allowing the conformational changes needed for 

infectivity. iv) In collaboration with J.L.Neira (CBMC) y M.A.Martínez (IRSI-Caixa), we are 

investigating the capacity of different synthetic peptides and other molecules to inhibit 

human immunodeficiency virus assembly and infectivity. These studies may be important 

for the design of new anti-HIV agents. v) In collaboration with G.Rivas (CIB), we have 

used two model viral systems to show that, in physiological macromolecular crowding 

conditions, the inhibitory activity of small-size antiviral compounds may be reduced. 

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Figure 1. Topographic image of an individual MVM 

particle in liquid, obtained in our laboratory by atomic force 

microscopy. Immediately after the image is obtained, the 

microscope is used to apply a force on the particle; this 

allows the determination of its mechanical elasticity. 

Figure 2. Location on the FMDV capsid 

structure (A) of a collecion of lethal 

mutations at the interfaces between 

pentameric subunits (B, colored white), and 

of the different compensatory mutations 

that restored infectivity (C, colored green 

or orange). 

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Mauricio García Mateu 

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Postdoctoral: 

Rebeca Pérez Fernández 


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Rebeca Bocanegra Rojo 

Milagros Castellanos Molina 

Inmaculada López Pérez 

Pablo José Pérez Carrillo 

Verónica Rincón Forero 


Miguel Ángel Fuertes Villadangos 

Alicia Rodriguez Huete 

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Carrasco, C., Douas, M., Miranda, R., Castellanos, M., Carrascosa, J.L., Mateu, M.G., Marqués, M. and 

de Pablo, P.J. (2009). Action of capillary forces of water confined at the nanoscale during dessication of 

viruses. Proc. Natl. Acad. Sci. USA 106, 5475-5480. (A) 

Serena, P.A., Douas, M., Marqués, M.I., Carrasco, C., de Pablo, P.J., Miranda, R., Carrascosa, J.L., 

Castellanos, M. and Mateu, M.G. (2009). MC simulations of water meniscus in nanocontainers: explaining 

the collapse of viral particles due to capillary forces. Phys. Status Solidi C 6, 2128-2132. 


Staff 

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Patents 

Mateu, M.G. (2009). Capsid protein interactions in human immunodeficiency virus type 1 assembly. FEBS 

J. 276, 6098-6109. 

Luna, E., Rodriguez-Huete, A., Rincón, V., Mateo, R. and Mateu, M.G. (2009). A systematic study of the 

genetic response of a variable virus to the introduction of deleterious mutations in a functional capsid 

region. J.Virol. 83, 10140-10151. 

Martín-Acebes, M., Rincón, V, Mateu, M.G. and Sobrino, F. (2010). A single amino acid substitution in the 

structural protein VP3 of foot-and-mouth disease virus can increase acid-lability and confer resistance to 

acid-dependent uncoating inhibition. J.Virol. 84, 2902-2912. 

Domenech, R., Abián, O., Bocanegra, R., Correa, J., Sousa-Herves, A., Riguera, R., Mateu, M.G., 

Fernández-Megía, E., Velázquez-Campoy, A. and Neira, J.L. (2010) Dendrimers bind the homodimerization 

interface of the capsid protein of HIV-1. Biomacromolecules 11, 2069-2078. 


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Mauricio G. Mateu, member of the Editorial Board of Virus Research 

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OEPM Patent nº 2 323 929 (Foot-and-mouth disease vaccine). Granted June 2010. 

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Eva Luna García (2010). Aproximaciones a la obtención de cápsidas más estables del 

virus de la fiebre aftosa y estudio de la generación de mutaciones compensatorias. 

Universidad Autónoma de Madrid. Director: Mauricio G. Mateu. 


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So-called mobile genetic elements (MGE), e.g. phages, plasmids, transposons and ICEs, 

can be transferred horizontally between cells and affect the genetic make-up and hence 

the behaviour of bacteria. Accordingly, horizontal gene transfer (HGT) has a crucial role in 

microbial evolution and has important implications in a myriad of environmental and public 

health problems. For instance, HGT is mainly responsible for the emergence and rapid 

dispersion of antibiotic resistance. 

Little is known, especially in Gram positive bacteria, about the mechanisms by which 

MGE exert their behavioural effects on their host or on regulation of their mobility. A better 

understanding of these issues is warranted to face important threats of for instance antibiotic 

resistance. In our laboratory we study these issues using as host Bacillus subtilis and we 

limit the MGE to plasmids and phages. 

We use B. subtilis because (i) it is probably the best studied Gram-positive bacterium; (ii) 

it is non-pathogenic; (iii) it is amenable to genetic manipulation due to its ability to develop 

natural competence; and (iv) B. subtilis is related to pathogenic/fastidious bacteria like 

Bacillus anthracis, B. cereus and, although more distantly, to Listeria monocytogenes. 

For some phages it has been described that they alter the behaviour of B. subtilis upon 

infection. However, neither sequence nor mechanistic information of how these phages 

affect behaviour of their infected host is available. We are studying these focussing on two 

phages. About 20% of the natural isolates of B. subtilis contain an endogenous plasmid. 

Most large plasmids (>10 kb) can be transferred to other cells via the process of conjugation. 

We are the first to have sequenced two large Bacillus plasmids and have identified genes 

involved in conjugation as well as others that probably affect behavioural processes of the 

host. We are now analyzing these to unravel regulation of the conjugation process and to 

gain insight in the way plasmid-located genes alter the life style of the host. 

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Figure 1. Phase contrast 

microscopy image of 

sporulating B. subtilis cells 

harbouring conjugative 

plasmid pLS20. 


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Figure 2. Genetic 

map of B. pumilus 

sporulation inhibiting 

plasmid p576. 

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Wilfried J.J. Meijer 

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Praveen K. Singh 

Gayetri Ramachandran 


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Esther Serrano 

Sandra Ballestero Beltrán 


Adriana Marulanda Aguirre 

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Singh, P. K., Ballestero-Beltrán, S., Ramachandran, G. and Meijer, W.J.J. (2010) Complete nucleotide 

sequence and determination of the replication region of the sporulation inhibiting plasmid p576 of Bacillus 

pumilus NRS576. Res. Microbiol. 161, 772-782. 

Muñoz-Espín, D., Daniel, R., Kawai, Y., Carballido-López, R., Castilla-Llorente, V., Errington, J.*, Meijer, 

W.J.J.*, and Salas, M.* *: contributed equally. (2009) The actin-like MreB cytoskeleton organizes viral DNA 

replication in bacteria. Proc. Natl. Acad. Sci. USA. July 27 Epub ahead of print. 


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Castilla-Llorente, V., Meijer, W.J.J., and Salas, M. Differential Spo0A-mediated effects on transcription and 

replication of the related Bacillus subtilis phages Nf and f29 explain their different behaviours in vivo. 

(2009) Nucleic Acids Res. 37: 4955-4964. 

Castilla-Llorente, V., Salas, M., and Meijer, W.J.J. (2009) Different responses to Spo0A-mediated suppression 

of the related Bacillus subtilis phages Nf and f29. Environ. Microbiol. 11: 1137-49. 

Publications 

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The human immunodeficiency virus (HIV) is a retrovirus that infects cells of the immune 

system, and is the etiological agent of AIDS. Nowadays, treatment of HIV-infection involves 

the use of inhibitors of retroviral enzymes such as the reverse transcriptase (RT), protease 

and integrase; as well as drugs targeting viral entry. Combination therapies including two 

or three RT inhibitors have been rather successful in the clinic, and play a key role in the 

management of HIV-infected individuals. However, the emergence of resistant viruses, 

the observed cross-reactivity between the inhibitors, and unwanted secondary effects are 

major hurdles towards their long-term efficacy. 

We are interested in therapeutic targets for HIV, emphasizing on the role of the viral 

RT. HIV RT plays a pivotal role in the replication of the viral genomic RNA. Recently, 

our efforts have been directed towards two major goals: (1) understanding the role of 

different amino acids in the nucleotide specificity of the enzyme, as well as in its fidelity of 

DNA synthesis; and (2) the elucidation of molecular mechanisms involved in RT inhibitor 

resistance and the analysis of the contribution of different amino acid substitutions in the 

acquisition of drug resistance. 

Nucleotide specificity studies are relevant in drug resistance but are also important to 

develop useful tools in molecular biology. Thus, we are trying to obtain more stable 

and faithful RTs, with biotechnological applications (for example, in the analysis of gene 

expression). Understanding the role of different residues in RT function (both on DNA 

polymerase activity, but also in modulating RNase H function and reverse transcription 

initiation) should help us to design novel strategies for treating HIV infection. 

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Figure 1. Crystal structure of HIV-1 reverse 

transcriptase complexed with an RNA/DNA 

template-primer, showing (in purple) thumb 

subdomain residues that modulate nucleoside 

analogue resistance 

Figure 2. ATP-mediated excision kinetics 

of AZT-monophosphate from blocked 

primer-template DNA/DNA complexes 

by wild-type and mutant HIV-1 reverse 

transcriptases. 

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Luis Menéndez Arias 


Mar Álvarez García 

Tania Matamoros Grande 


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Verónica Barrioluengo Fernández 

Gilberto J. Betancor Quintana 

Mónica Kisic Aguirre 


Raquel N. Afonso Lehmann 

Daniela Barbieri 

Yasemin Ezgi Ertürk 

Luis Gosálbez Cisneros-Miret 

Cristina Jiménez Sánchez 

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Publications 

Álvarez, M., Matamoros, T. and Menéndez-Arias, L. (2009) Increased thermostability and fidelity of DNA 

synthesis of wild-type and mutant HIV-1 group O reverse transcriptases. J. Mol. Biol. 392, 872-884. 

De Mendoza, C., Anta, L., García, F., Pérez-Elías, M.J., Gutiérrez, F., Llibre, J.M., Menéndez-Arias, L., 

Dalmau, D., Soriano, V., on behalf of Platform for Drug Resistance of the Spanish AIDS Research Network 

(2009) HIV-1 genotypic drug resistance interpretation rules – 2009 Spanish guidelines. AIDS Rev., 11, 39-51. 

Garriga, C., Pérez-Elías, M.J., Delgado, R., Ruiz, L., Pérez-Álvarez, L., Pumarola, T., López-Lirola, A., 

González-García, J. and Menéndez-Arias, L., on behalf of the Spanish Group for the Study of Antiretroviral 

Drug Resistance (2009) HIV-1 reverse transcriptase thumb subdomain polymorphisms associated with 

virological failure to nucleoside drug combinations. J. Antimicrob. Chemother. 64, 251-258. 

Matamoros, T., Nevot, M., Martínez, M.A. and Menéndez-Arias, L. (2009) Thymidine analogue resistance 

suppression by V75I of HIV-1 reverse transcriptase: Effects of substituting Valine 75 on stavudine excision 

and discrimination. J. Biol. Chem. 284, 32792-32802. 

Menéndez-Arias, L. (2009) Mutation rates and intrinsic fidelity of retroviral reverse transcriptases. Viruses 

1, 1137-1165; doi:10.3390/v1031137 

Puertas, M.C., Buzón, M.J., Artese, A., Alcaro, S., Menéndez-Arias, L., Perno, C.F., Clotet, B., Ceccherini- 

Silberstein, F. and Martinez-Picado, J. (2009) Effect of the human immunodeficiency virus type 1 reverse 

transcriptase polymorphism Leu-214 on replication capacity and drug susceptibility. J. Virol. 83, 7434-7439. 

Betancor, G., Puertas, M.C., Nevot, M., Garriga, C., Martínez, M.A., Martinez-Picado, J. and Menéndez- 

Arias, L. (2010) Mechanisms involved in the selection of HIV-1 reverse transcriptase thumb subdomain 

polymorphisms associated with nucleoside analogue therapy failure. Antimicrob. Agents Chemother. 

54, 4799-4811. 

Clotet, B., Menéndez-Arias, L., Schapiro, J. M., Kuritzkes, D., Burger, D., Telenti, A., Brun-Vezinet, F., Geretti, 

A. M., Boucher, C. A. and Richman, D. D. (eds.) (2010) Guide to management of HIV drug resistance, 

antiretrovirals pharmacokinetics and viral hepatitis in HIV infected subjects, 10th Edition, Fundació de Lluita 

contra la SIDA, Barcelona, Spain, 422 pp. 

Menéndez-Arias, L. (2010) Special issue: Retroviral enzymes. Viruses 2, 1181-1184 (editorial). 

Menéndez-Arias, L. (2010) Molecular basis of human immunodeficiency virus drug resistance: An update. 

Antiviral Res. 85, 210-231. 

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Antirretroviral / human immunodeficiency virus reverse transcriptase and antire- 

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Member of the Editorial Boards of Antiviral Therapy, Viruses and Timely Topics in Medicine: 

AIDS Cyber Journal. 

Editor of a special issue of the on-line journal Viruses dedicated to “Retroviral enzymes” 

(2009/2010). 


Member of the formation and training panel of the Spanish AIDS Research Network, 

and co-organizer of its “Third Training Symposium” (San Lorenzo de El Escorial, 

October 7-9, 2009). 

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L. Menéndez Arias, T. Matamoros, M. Álvarez (2010). Retrotranscriptasa del VIH-1 de 

grupo O modificada. Ref.: PCT/ES2010/070320. 


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Mónica Kisic Aguirre (2010). Mecanismos moleculares de resistencia e hipersensibilidad 

a fármacos antirretrovirales mediados por deleciones en la horquilla β3-β4 de la retrotranscriptasa 

del virus de la inmunodeficiencia humana tipo 1. Universidad Autónoma 

de Madrid. Director: Luis Menéndez Arias. 


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Repair of African swine fever virus (ASFV) DNA. 

Presently, the objective of this work is to study the biological function of the reparative DNA 

polymerase, pol X, of ASFV, which participates, together with the viral AP endonuclease 

and DNA ligase, in a base excision repair (BER) pathway. Our studies show that pol X is 

required for virus replication in its natural host cell, the swine macrophage, and protects 

the infected cell against oxidizing genotoxic agents. Moreover, pol X is necessary to 

maintain genome stability, by repairing mutations introduced in the viral DNA during the 

infection, which underlines the importance of its in vivo function. 

ASFV morphogenesis. 

The biogenesis of ASFV membranes is one of the most interesting and controversial 

subjects in the study of virus morphogenesis. In order to undertake these studies, we 

are using ASFV recombinants that inducibly express membrane structural proteins 

localized in the inner envelope of the virus particle.We have recently identified a new 

morphogenetic intermediate constituted by membrane helicoidal structures that are 

precursors of icosahedral viral particles. To get a better knowledge of the structure of these 

helicoidal intermediates, we are presently using electron tomography and other advanced 

techniques in electron microscopy for the resolution of three-dimensional structures. 

Another objective of our studies on virus morphogenesis is to understand the regulation of 

ASFV polyproteins pp220 and pp62 processing. The polyproteins are major components 

of the viral core, being their proteolytic processing essential for virus maturation. Our 

results indicate the existence of redox mechanisms that might operate during the infective 

cycle by regulating the activity of the viral protease that catalyzes the processing of 

polyproteins. 

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Figure 1. Serial section analysis of a viral factory showing a helicoidal membrane structure 

with an associated icosahedral particle. Right panel, reconstruction of the helicoidal structure. 

The components of each strand in the different sections are identically colored. 

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Maria Luisa Salas Falgueras 

Postdoctoral: 

Modesto Redrejo Rodríguez 

Cristina Suárez García 


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Cristina Suárez García 

Marina del Rosal Macías 


María Luisa Nogal París 

María José Bustos Sánchez 

Esther Martín Forero 


María Ballester Devis 

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Redrejo-Rodríguez, M., Ishchenko, A. A., Saparbaev, M. K., Salas, M. L. and Salas, J. (2009) African swine 

fever virus AP endonuclease is a redox-sensitive enzyme that repairs alkylating and oxidative damage to 

DNA. Virology 390, 102-109. 

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Rodríguez, I., Nogal, M. L.,Redrejo-Rodríguez, M., Bustos, M. J.and Salas, M. L. (2009) The African swine 

fever virus virion membrane protein pE248R is required for virus infectivity and an early postentry event. J. 

Virol. 83, 12290-12300. 


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Publications 

Suárez, C., Salas, M. L. and Rodríguez, J. M. (2010) African swine fever virus polyprotein pp62 is essential 

for viral core development. J. Virol. 84, 176-187. 

Suárez, C., Gutiérrez-Berzal, J., Andrés, G., Salas, M. L. and Rodríguez, J. M. (2010) The African swine 

fever virus protein p17 is essential for the progression of the viral membrane precursors towards icosahedral 

intermediates. J. Virol. 84, 7484-7499. 

Ballester, M., Galindo-Cardiel, I., Gallardo, C., Argilaguet, J. M., Segalés, J., Rodríguez, J. M. and Rodríguez, 

F. (2010) Intranuclear detection of African swine fever virus DNA in several cell types from formalin-fixed and 

paraffin-embedded tissues using a new in situ hybridization protocol. J. Virol. Methods 168, 38-43. 

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M. L. Salas y J. M. Rodríguez (CSIC), L. K. Dixon (IAH). (2010). Vacuna contra el virus 

de la peste porcina Africana basada en virus recombinantes deficientes en replicación. 

Nº Solicitud: ES1641.748. 


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Modesto Redrejo Rodríguez (2009). La endonucleasa AP del virus de la peste porcina 

Africana y el papel biológico del sistema viral de reparación del ADN. Universidad 

Autónoma de Madrid. Directores: José Salas Falgueras y María Luisa Salas Falgueras. 

Cristina Suárez García (2009). Papel de la poliproteína pp62 y de la proteína p17 en la 

morfogénesis del virus de la peste porcina Africana. Universidad Autónoma de Madrid. 

Director: Javier M. Rodríguez. 


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New strategies for prevention and control of viral diseases: foot-and-mouth 

disease virus as a model 

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Foot-and-mouth disease virus (FMDV) is one of the major concerns for animal health. It 

is also an interesting model system for understanding the interactions of a highly variable 

virus and its natural hosts and the implications of these interactions on disease control. 

We are working in the development of new FMDV marker vaccines (peptides and DNA 

vaccines) that can induce protective humoral and cellular immune responses, using as 

animal model an important natural host: the pig. Some of these strategies are also being 

applied to the development of new vaccines against classical swine fever (CSF). We are 

also analyzing the functional role of FMDV proteins on the internalization, the replication 

cycle and the mechanisms mediating the pathogenesis of FMDV and other related viruses 

causing vesicular diseases, such as swine vesicular disease virus (SVDV), and vesicular 

stomatitis virus (VSV) in cultured cells and in animal models. Special attention is being 

paid to the functional implications of non-structural proteins, like those from the FMDV 

3AB region, in virus virulence and host range. A parallel study of the functional implications 

of non-coding RNA regions is also being conducted. Besides providing basic information 

on the multiplication cycle of these viruses, the results obtained are being used for the 

identification of antiviral targets, attenuation determinants as well as the design of new 

vaccine strategies. As part of these studies, we are characterizing the recently identified 

inhibitory effect of valproic acid on the multiplication of enveloped viruses. 

Part of the work has been carried out in the BSL3 facilities at CISA-INIA. 

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Figure 1. FMDV particles (red) entering IBRS-2 

cells. Nuclei stained in blue. 

Figure 2. Colocalization assays in IBRS-2 cells of 

SVDV (blue), transferrin (green) and tubulin (red). 

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Francisco Sobrino 

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Margarita Sáiz 

Postdoctoral: 

Maria Flora Rosas 


Raúl Postigo 

Miguel Ángel Martín 

María Teresa Sánchez 

Miguel Rodríguez 

Yuri A. Vieira 

Ángela Vázquez 

Flavia Caridi 


Susan Realgen 

Abraham Arrizon 

Maria Rodolis 

Jackob Zimmermann 


Mónica González 

Visiting Scientist: 

Belén Borrego (CISA-INIA) 


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Rodríguez-Pulido, M., Sobrino, F., Borrego, B., Sáiz, M. (2009) RNA. Attenuated foot-and-mouth disease virus 

RNA carrying a deletion in the 3´ non-coding region can elicit immunity in swine J. Virol.. 83, 3475-3485. 

Martín-Acebes, M.A., González-Magaldi, M., Vázquez-Calvo, A., Armas-Portela, R. and Sobrino, F. (2009) 

Internalization of swine vesicular disease virus into cultured cells: a comparative study with foot-and-mouth 

disease virus. J. Virol. 83, 4216-4266 


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Publications 

Patents 

Sanchez-Aparicio, M.T., Rosas,M-F., Ferraz, R.M., Delgui, L., Veloso, J.J., Blanco, E, Villaverde, A. and 

Sobrino, F. (2009) Discriminating foot-and-mouth disease virus-infected and vaccinated animals by use of 

β-galactosidase allosteric biosensors. Clin. Vaccine Immunol.16, 1228-1235 

Martín-Acebes, MA., Rincón, V., Armas-Portela, R., Mateu, M.G. and Sobrino, F. (2010). A single amino acid 

substitution in the capsid of foot-and-mouth disease virus can increase acid-lability and confer resistance 

to acid-dependent uncoating inhibition. J. Virol. 84, 2902-2912 

Rodríguez-Pulido, M., Sobrino, F., Borrego, B. and Sáiz, M. (2010). RNA immunization can protect mice 

against foot-and-mouth disease virus. Antiviral Res. 85, 556–558. 

Martín-Acebes, M.A., Herrera, M., Domingo, E. and Sobrino, F. (2010). Cell density-dependent expression 

of viral antigens during persistence of foot-and-mouth disease virus in cell culture. Virology 403, 47–55. 


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Construcción peptídica dendrimérica para la prevención de la fiebre aftosa en animales 

Inventores: C. Cubillo, E. Blanco, J. Bárcena, F. Sobrino y D. Andreu. Nº de solicitud: 

P 200602142. Fecha de prioridad: 2-08-2006. Solicitud PCT (08/2007) Nº PCT ES 

2007/070146. 

Uso del ácido valproico como antiviral contra virus con envoltura. A. Vazquez, F. Sobrino, 

M.A. Martín y J.C. Sáiz. No Solicitud: P 200602142 (29 marzo 2010). 


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Miguel Ángel Martín Acebes (2009). Mecanismos de entrada y de organización del 

complejo de replicación del virus de la fiebre aftosa: estudio comparativo con el virus de 

la enfermedad vesicular del cerdo y el virus de la estomatitis vesicular. Universidad Autónoma 

de Madrid. Co-dirigida por F. Sobrino y R. Armas. 


Staff 

Publications 

Patents 

Miguel Ramón Rodríguez Pulido. Julio 2009. Estructura y función de la región 3´ no codificante 

del virus de la fiebre aftosa. Aplicación a nuevas estrategias vacunales basadas 

en RNA. Universidad Autónoma de Madrid. Director: M. Sáiz. 

Raúl Postigo Fernández. Octubre 2009. Análisis funcional de la proteína 3A del virus de 

la fiebre aftosa. Universidad Autónoma de Madrid. Co-dirigida por F. Sobrino y M.F. Rosas. 

María Teresa Sánchez Aparicio. Abril 2010. Estudio de proteínas no estructurales del 

virus de la fiebre aftosa: análisis funcionales y aplicación al diagnóstico viral. Co-dirigida 

por F. Sobrino y M.F. Rosas. Universidad Autónoma de Madrid. 


CBMSO 2009-2010

03B Virology and Microbiology.pdf - Universidad AutÃ³noma de Madrid

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