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The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Session 11 abstracts 64-67: Gene Expression II 92
The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Gene Expression II Abstract 64 DEVELOPMENT OF A COMPREHENSIVE TRANSCRIPTIONAL NETWORK FOR KSHV Linda Persson, Scott Millman and Angus Wilson Department of Microbiology & NYU Cancer Institute, New York University School of Medicine, New York, NY 10016, USA Abstract Viral transcription factor RTA (product of ORF50) plays an essential and far-reaching role in control of KSHV replication and pathogenesis. In latently infected cells, the silent ORF50 promoter responds to a variety of reactivation stimuli, leading to rapid accumulation of RTA protein and initiation of the lytic program. A principal function of RTA is to direct the transcription of more than 80 genes required for the lytic replication and virion assembly. Several RTA targets encode positive and negative regulators which further mold the basic program into a cascade of sequential viral gene expression. Paradoxically, RTA may also contribute to the establishment of latency during de novo infection, acting through an inducible promoter (LTi) upstream of the K12-ORF73 latency gene cluster. To better understand the intricate network of regulatory interactions that control lytic replication and determine how the program can be modified by environmental parameters, we embarked on a comprehensive analysis of the RTAdirected transcriptome. We began with a functional screen to identify RTA-responsive sequences irrespective of their position in the genome. A pilot study covering 37,300-bp between K8.1 and ORF66 revealed 8 separate RTA-responsive elements, corresponding to a mix of new and known lytic promoters. Currently, this unbiased mapping approach is being applied to the entire KSHV genome. Mutational analysis and bioinformatics methods are being used to dissect the individual response elements and identify points at which cellular signaling pathways intersect with RTA to fine tune the lytic program or express only a subset of viral genes. Presenting author Email: wilsoa02@med.nyu.edu 93
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