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The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Poster Session Abstract P13 IDENTIFICATION OF NOVEL CONSERVED GAMMAHERPESVIRUS MICRORNAS Nicole Walz , Thomas Christalla and Adam Grundhoff Heinrich-Pette Institute for Experimental Virology and Immunology, D-20251 Hamburg, Germany Abstract microRNAs (miRNAs) are small (~22 nt.), non-coding RNA molecules which posttranscriptionally regulate mRNA expression and play important roles in the regulation of diverse cellular processes. The current release of the miRNA database (miRBase) lists a total of 132 known viral miRNAs, the vast majority of which are encoded by members of the herpesvirus family. While the propensity to encode miRNAs appears to be a conserved feature of many herpesviruses, the miRNAs themselves show little conservation: with the exception of 7 miRNAs known to be shared between Epstein-Barr Virus (EBV) and its close relative Rhesus Lymphocryptovirus (rLCV), all other herpesvirus miRNAs appear unrelated in sequence. We have recently developped VMir, a computational ab initio prediction method to identify viral miRNAs, and have employed the program to identify miRNAs in SV40, KSHV and EBV. We have modified the algorithm to allow consideration of evolutionary conservation and conducted an extended analysis of the herpesvirus family. We were able to identify and experimentally verify 2 novel EBV- and 17 novel rLCV-encoded miRNAs; with a total of 33 miRNAs rLCV is now the virus with the highest number of known miRNAs. In contrast, we have found little evidence for the existence of hitherto unknown conserved miRNAs shared by other herpesviruses, including KSHV and the closely related Rhesus Rhadinovirus (RRV). While EBV and rLCV thus share even more miRNAs than previously thought, our results underline the notion that, in contrast to their cellular counterparts, viral miRNAs show an almost total lack of sequence conservation. Presenting author Email: Adam.Grundhoff@hpi.uni-hamburg.de 110
The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Poster Session Abstract P14 LYTIC INDUCTION OF KAPOSI'S SARCOMA-ASSOCIATED HERPESVIRUS IN PRIMARY EFFUSION LYMPHOMA CELLS WITH NON-TOXIC NATURAL PRODUCTS Hye-Jeong Cho 1 , Fuqu Yu 2 , Ren Sun 2 , Dongho Lee 1 and Moon Jung Song 1 1 Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 136-713, Republic of Korea, and 2 Department of Molecular and Medical Pharmacology, University of California at Los Angeles, Los Angeles, CA90095, U.S.A. Abstract Kaposi’s sarcoma-associated herpesvirus (KSHV) has been linked to Kaposi’s sarcoma, primary effusion lymphoma (PEL), and multicentric Castleman’s disease. Intentional lytic induction of gammaherpesviruses in the presence of antiviral drugs is thought to be an effective treatment option for gammaherpesvirus-related tumors. In this study, we used a cell-based fluorescence bioassay system in which a KSHV-infected PEL cell line was stably transfected with a potent viral promoter-driven reporter gene to identify effective non-toxic reagents capable of inducing latent KSHV. Among 400 plant extracts screened, three extracts increased reporter gene expression in a dose-dependent manner. The three extracts activated the RTA promoter and induced expressions of lytic genes in the endogenous viral genomes of KSHV-infected tumor cells. Furthermore, these extracts were also capable of inducing lytic replication of Epstein-Barr virus in B95.8 cells, suggesting a conserved reactivation mechanism(s) among gammaherpesviruses. Together, our results demonstrate the effectiveness of the screening system to identify natural products capable of inducing KSHV reactivation, thereby facilitating the development of novel therapeutic agents for KSHV-associated malignancies. Presenting author Email: chohyejeong@korea.ac.kr 111
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