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The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK
Latency Abstract 1
DELETION OF LANA FROM RHESUS RHADINOVIRUS (RRV) GENERATES A
HIGHLY LYTIC RECOMBINANT VIRUS
Kwun Wah Wen, Chelsey Hilscher, Dirk P. Dittmer and Blossom Damania
Department of Microbiology and Immunology and Lineberger Cancer Center, University of
North Carolina at Chapel Hill, North Carolina 27599.
Abstract
Rhesus monkey rhadinovirus (RRV) is a gamma herpesvirus that is closely related to the
human Kaposi's sarcoma-associated herpesvirus (KSHV/HHV-8). RRV serves as an in
vitro and an in vivo model for KSHV. RRV can be grown to high titers on rhesus
fibroblasts and the availability of the RRV lytic system facilitates analysis of viral
replication and the contribution of individual open-reading frames to viral fitness. We
had previously reported that RRV LANA (R-LANA) can suppress lytic viral replication and
that R-LANA inhibits Rta/Orf50 transactivation of lytic promoters resulting in lower viral
titers (Dewire and Damania, J. Virology, 2005). Here we present data on the construction
of a RRVΔLANA/GFP recombinant virus by homologous recombination. Integrity of the
recombinant virus was confirmed by Southern blot analysis, restriction digest and PCR.
We compared replication kinetics of RRVΔLANA/GFP, RRV-GFP, wild-type (WT) RRV, and
a revertant virus (RRVREV) using one-step growth curves. Viral replication was quantitated
using traditional plaque assays as well as real-time PCR based genome quantification
assay. We found that the RRVΔLANA/GFP recombinant virus exhibits highly lytic
replicative properties compared to RRV-GFP, WT RRV, and the revertant virus. We also
employed a quantitative real time PCR-based RRV viral array to transcriptionally profile
lytic gene expression during de novo infection using RRVΔLANA/GFP and RRV-GFP
recombinant viruses. The RRVΔLANA/GFP virus displayed increased lytic gene expression
at all time points post-infection compared to RRV-GFP.
Presenting author E-mail: kenwen@med.unc.edu
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