# Mechanisms of HIV-1 RNA Methylation in Regulating Viral Replication

> **NIH NIH R01** · UNIVERSITY OF IOWA · 2020 · $383,356

## Abstract

Project Summary/Abstract
Our goal is to investigate the functions and mechanisms of N6-methyladenosine (m6A) modification of
HIV-1 RNA during viral infection. The internal m6A modification of cellular RNAs is a novel mechanism
of post-transcriptional control of gene expression, which is coordinately regulated by three groups of
host proteins, including methyltransferases (writers), demethylases (erasers), and m6A-selective-
binding proteins (readers). Binding of m6A-modified cellular RNA by the readers significantly affects
various aspects of RNA functions during translation. Three recent publications, including one from our
group, highlighted the critical role of m6A modification of HIV-1 RNA in regulating viral replication.
However, these studies reported some different results and suggested distinct mechanisms. To clarify
the discrepancy, it is essential to define the mechanisms by which m6A modification of HIV-1 RNA
regulates viral replication in primary CD4+ T-cells.
We identified multiple regions of m6A modification in HIV-1 RNA bound by the readers in HIV-1-
infected primary CD4+ T-cells. We found that the expression levels of the readers significantly
modulated HIV-1 infection in target cells. Knockdown of the writers or erasers in virus-producing cells
significantly affected HIV-1 Gag protein synthesis, suggesting the importance of m6A modification of
HIV-1 RNA in regulating viral gene expression. Interestingly, we observed that HIV-1 infection
upregulated the expression of endogenous readers in primary CD4+ T-cells. Our central
hypothesize is that reversible m6A modification of HIV-1 RNA regulates viral replication in CD4+ T-
cells by affecting the structure, stability, splicing, and/or trafficking of HIV-1 RNA. We will test this
hypothesis in three specific aims using interdisciplinary approaches. Aim 1. To identify the m6A
residues in HIV-1 RNA and to investigate the effects of m6A modification on HIV-1 RNA structure and
interactions with the reader proteins; Aim 2. To investigate the effects of m6A modification on the
stability, splicing and trafficking of HIV-1 RNA in infected primary CD4+ T-cells; and Aim 3. To
investigate the effects of HIV-1 infection and proinflammatory cytokines on the expression and
localization of the readers, writers, and erasers and to define the underlying mechanisms.
Overall impact: Accomplishing our proposed studies will define the mechanisms by which HIV-1 RNA
m6A modification regulates viral infection in CD4+ T-cells. Investigations of the HIV-1 RNA m6A
modification and interactions with host proteins represent a new area of HIV-1 RNA biology, which
can facilitate therapeutic development against HIV-1 infection.

## Key facts

- **NIH application ID:** 9961494
- **Project number:** 5R01AI150343-05
- **Recipient organization:** UNIVERSITY OF IOWA
- **Principal Investigator:** Li Wu
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $383,356
- **Award type:** 5
- **Project period:** 2019-11-01 → 2022-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9961494

## Citation

> US National Institutes of Health, RePORTER application 9961494, Mechanisms of HIV-1 RNA Methylation in Regulating Viral Replication (5R01AI150343-05). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/9961494. Licensed CC0.

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