# Regulation of KSHV replication by N6-methyladenosine (m6A)

> **NIH NIH R01** · UNIVERSITY OF PITTSBURGH AT PITTSBURGH · 2022 · $339,863

## Abstract

N6-methyladenosine (m6A) is the most abundant internal modification on poly(A) RNA. Dynamic regulation of
the m6A epitranscriptome is involved in diverse cellular functions. m6A mediates these functions by affecting
mRNA translation, alternative splicing, nuclear export and degradation, and miRNA biogenesis and binding
through three groups of proteins: methyltransferases or “writers”, demethylases or “erasers”, and m6A-binding
proteins or “readers”. m6A is present in the genomes of RNA viruses and regulates the replication of these
viruses. Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiologic agent of Kaposi's sarcoma (KS) and
primary effusion lymphoma (PEL) commonly found in AIDS patients. Understanding the mechanism regulating
KSHV latent and lytic replication can not only provide insights into the pathogenesis of KSHV-induced cancers
but also serve as the basis for developing novel therapy. In the current funding period, we have made
significant progresses toward this goal. For this renewal, we have discovered that m6A is abundant on KSHV
transcripts, and that m6A reader protein YTHDF2 acts as an antiviral factor during viral lytic replication. We
demonstrate m6A dynamics during KSHV latent and lytic replications, and in different cell types that support
distinct viral replication programs. Despite these works, the roles of m6A in KSHV infection have just started to
be revealed. The Objective of this application is to systematically map the dynamics of m6A modifications at a
single base resolution and bindings of m6A reader proteins in KSHV epitranscriptome, and determine their
functions in different phases of KSHV life cycle, and KSHV-induced tumorigenesis. The Central Hypothesis is
that KSHV m6A modifications are dynamically regulated, and these modifications mediate different phases of
KSHV life cycle, and hence KSHV-induced tumorigenesis. We will test this hypothesis by mapping m6A marks
in KSHV transcriptome at a single base resolution, and determine the roles of m6A writer and eraser proteins in
different phases of KSHV life cycle and KSHV-induced tumorigenesis (Aim 1); determining the functions of
m6A reader proteins in different phases of KSHV life cycle by gain- and loss-of-function approaches and by
examining their bindings to KSHV transcripts (Aim 2); and examining the functions of KSHV m6A marks in the
context of viral infection using Crispr-Cas9-guided m6A writer and eraser, and by site-specific mutagenesis
(Aim 3). It is our expectations that this project will provide comprehensive mapping and functional delineation
of m6A marks, and m6A writer, eraser and reader proteins in different phases of KSHV life cycle, and KSHV-
induced tumorigenesis. This work is highly significant as it will, for the first time, systematically reveal the
functions of these RNA modifications in KSHV infection, thus providing insights into the mechanism regulating
KSHV life cycle and KSHV-induced pathogenesis. This study will also identify pot...

## Key facts

- **NIH application ID:** 10466778
- **Project number:** 5R01CA124332-14
- **Recipient organization:** UNIVERSITY OF PITTSBURGH AT PITTSBURGH
- **Principal Investigator:** Shou-Jiang Gao
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $339,863
- **Award type:** 5
- **Project period:** 2007-04-01 → 2026-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10466778

## Citation

> US National Institutes of Health, RePORTER application 10466778, Regulation of KSHV replication by N6-methyladenosine (m6A) (5R01CA124332-14). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10466778. Licensed CC0.

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