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The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Latency Abstract 6 KSHV ENCODED LANA INTERACTS WITH THE NUCLEAR MITOTIC APPARATUS PROTEIN TO REGULATE GENOME MAINTENANCE AND SEGREGATION Huaxin Si 1 , Subhash C. Verma 1 , Michael Lampson 2 , Qiliang Cai 1 , Erle S. Robertson 1 1Department of Microbiology and the Tumor Virology Program of the Abramson Comprehensive Cancer Center, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania, 19104 2 Department of Biology, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania, 19104 Abstract Kaposi’s sarcoma associated herpesvirus (KSHV) genomes are tethered to the host chromosomes and partitioned faithfully into daughter cells with the host chromosomes. The latency associated nuclear antigen (LANA) is important for segregation of the newly synthesized viral genomes to the daughter nuclei. Here we report that the Nuclear Mitotic Apparatus protein (NuMA) and LANA can associate in KSHV infected cells. In synchronized cells, NuMA and LANA are colocalized in interphase cells and separate during mitosis at the beginning of prophase, reassociating again at the end telophase and cytokinesis. Silencing of NuMA expression by siRNA and expression of LGN and a dominant negative of dynactin (P150-CC1) which disrupts the association of NuMA with microtubule resulted in the loss of KSHV terminal repeats plasmids containing the major latent origin. Thus, NuMA is required for persistence of the KSHV episomes to daughter cells. This interaction between NuMA and LANA is critical for segregation and maintenance of the KSHV episomes through a temporally controlled mechanism of binding and release during specific phases of mitosis. Presenting author Email: vermas@mail.med.upenn.edu 26
The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Latency Abstract 7 MULTIPLE DOMAINS CONTRIBUTE TO THE ASSOCIATION OF LANA WITH HOST CHROMATIN COMPONENTS Ryo Nasu, Satoko Matsumura, Naoko Tanese and Angus Wilson Department of Microbiology & NYU Cancer Institute, New York University School of Medicine, New York, NY 10016, USA Abstract The latency-associated nuclear antigen (LANA, LANA1, LNA1) encoded by ORF73 is expressed in all KSHV infected cells and performs a variety of functions associated with maintenance of the viral genome and proliferation of the infected host cell. As its name indicates, LANA accumulates in the nucleus where it is associated with host chromosomes throughout the cell cycle. Direct interactions with a number of chromatin components have been described but the functional significance of these interactions is not fully understood. We have focused on the association of LANA with methyl-CpG binding protein 2 (MeCP2) and core histones. Evidence will be presented showing that both the N- and C-terminal domains of LANA contribute to interaction with MeCP2 and this can direct LANA to regions of pericentric heterochromatin. Localization is dependent on the methyl-CpG binding (MBD) and adjacent trans-repression (TRD) domains of MeCP2. The chromatin binding motif (CBM) at the extreme N-terminus of LANA (residues 5-11) influences subcellular localization but is not strictly required for MeCP2 binding. Instead, the CBM recognizes the four histones that form the nucleosome core. In addition to interactions with histones H2A/H2B previously described by the Kaye and Luger laboratories, we will present evidence for additional interactions with histones H3 and H4. Sedimentation studies employing recombinant proteins show that the LANA CBM is capable of assembling isolated nucleosomes into compacted structures that may serve to regulate host and viral gene expression. Presenting author Email: wilsoa02@med.nyu.edu 27
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Keiji Ueda Dept.of Infectious Disea
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Prof Juergen Haas Max-von-Pettenkof
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Vladimir Majerciak 4817 36th Street
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