# Epigenetic Regulation of KSHV Genome Replication

> **NIH NIH R01** · UNIVERSITY OF PENNSYLVANIA · 2022 · $501,823

## Abstract

Kaposi's sarcoma Associated Herpesvirus (KSHV) or Human Herpesvirus 8 (HHV-8) is an
oncogenic gammaherpesvirus known to be the causative agent of Kaposi's sarcoma (KS), and
contributes to body cavity based lymphomas (BCBLs) or pleural effusion lymphomas (PELs) in
AIDS patients. It is also associated with Multicentric Castleman's Disease (MCD). KSHV infects
endothelial and human B cells with expression of a limited repertoire of genes that are linked to
latent infection including the major latency associated nuclear antigen (LANA). KSHV
undergoes two major replication modes; a lytic mode and a latent replication mode and in some
instances there is an underlying low level of lytic replication that is seen during latency. This
may be critical for the pathogenesis associated with the virus. KSHV latent replication is
dependent on expression of LANA and initiates at the terminal repeats (TRs). LANA binding
sites have been mapped to the TR elements and these sites recruit replication proteins ORCs
and MCMs. We have shown that additional sites on the KSHV genome can initiate replication at
other regions shown to also recruit ORC and MCMs. A unique technology referred to as single
molecule analysis of replicated DNA (SMARD) was used to identify other regions capable of
incorporating fluorescent nucleoside analogs during cell proliferation. We have also shown that
the replication initiation zone is independent of the presence of LANA demonstrating that the
KSHV genome is capable of initiating replication during latency at multiple sites along the
genome. In this proposed application we will focus our efforts on understanding genome
replication of the KSHV virus after de novo infection by focusing on the major regions of the
genome that are activated for replication on infection of primary cells. We will determine the
epigenetic programming of the genome, and higher order conformations which dictates genome
sites containing firing capabilities for successful replication of the genome. Infected cells will be
harvested at different time points of infection and the replication zones monitored by SMARD.
We will compare these zones after infection of primary B- and endothelial cells. We will also
quantitate the semi-conservative replication using a Meselson Stahl modified approach with
real-time PCR. ChIP/ChIP-Seq and ChIA-PET-sequencing will be used to identify the genome
regions associated with replication proteins ORCs, MCMs, chromatin modifying factors, and
viral antigens. The analysis will determine the time points after the viral genome enters the
nucleus to obtain a temporal picture of the transitional epigenetic marks that are determinants
for replication. Furthermore, we will monitor the long range interactions, and conformation
changes that occur on the viral genome during de novo infection to understand the contribution
of epigenetics, higher order interactions and the viral and cellular antigens required for
replication of the KSHV genome after de n...

## Key facts

- **NIH application ID:** 10457380
- **Project number:** 5R01CA244074-04
- **Recipient organization:** UNIVERSITY OF PENNSYLVANIA
- **Principal Investigator:** ERLE S. ROBERTSON
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $501,823
- **Award type:** 5
- **Project period:** 2019-08-01 → 2024-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10457380

## Citation

> US National Institutes of Health, RePORTER application 10457380, Epigenetic Regulation of KSHV Genome Replication (5R01CA244074-04). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10457380. Licensed CC0.

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