Visualizing the translation and packaging of HIV-1 full-length RNA

Jianbo Chen, Yang Liu, Bin Wu, Olga A. Nikolaitchik, Preeti R. Mohan, Jiji Chen, Vinay K. Pathak, Wei Shau Hu

Research output: Contribution to journalArticlepeer-review

Abstract

HIV-1 full-length RNA (HIV-1 RNA) plays a central role in viral replication, serving as a template for Gag/Gag-Pol translation and as a genome for the progeny virion. To gain a better understanding of the regulatory mechanisms of HIV-1 replication, we adapted a recently described system to visualize and track translation from individual HIV-1 RNA molecules in living cells. We found that, on average, half of the cytoplasmic HIV-1 RNAs are being actively translated at a given time. Furthermore, translating and nontranslating RNAs are well mixed in the cytoplasm; thus, Gag biogenesis occurs throughout the cytoplasm without being constrained to particular subcellular locations. Gag is an RNA binding protein that selects and packages HIV-1 RNA during virus assembly. A longstanding question in HIV-1 gene expression is whether Gag modulates HIV-1 RNA translation. We observed that despite its RNA-binding ability, Gag expression does not alter the proportion of translating HIV-1 RNA. Using single-molecule tracking, we found that both translating and nontranslating RNAs exhibit dynamic cytoplasmic movement and can reach the plasma membrane, the major HIV-1 assembly site. However, Gag selectively packages nontranslating RNA into the assembly complex. These studies illustrate that although HIV-1 RNA serves two functions, as a translation template and as a viral genome, individual RNA molecules carry out only one function at a time. These studies shed light on previously unknown aspects of HIV-1 gene expression and regulation.

Original languageEnglish (US)
Pages (from-to)6145-6155
Number of pages11
JournalProceedings of the National Academy of Sciences of the United States of America
Volume117
Issue number11
DOIs
StatePublished - Mar 17 2020

Keywords

  • Gaggenome packaging
  • HIV
  • RNA
  • genome packaging
  • translation

ASJC Scopus subject areas

  • General

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