Suppression of HIV-1 Infection by APOBEC3 Proteins in Primary Human CD4(+) T Cells Is Associated with Inhibition of Processive Reverse Transcription as Well as Excessive Cytidine Deamination

Kieran Gillick, Darja Bamming, Prabhjeet Phalora, Eun-Young Kim, Steven M Wolinsky, Michael H Malim

Research output: Contribution to journalArticlepeer-review

88 Citations (Scopus)

Abstract

The Vif protein of human immunodeficiency virus type 1 (HIV-1) promotes viral replication by downregulation of the cell-encoded, antiviral APOBEC3 proteins. These proteins exert their suppressive effects through the inhibition of viral reverse transcription as well as the induction of cytidine deamination within nascent viral cDNA. Importantly, these two effects have not been characterized in detail in human CD4(+) T cells, leading to controversies over their possible contributions to viral inhibition in the natural cell targets of HIV-1 replication. Here we use wild-type and Vif-deficient viruses derived from the CD4(+) T cells of multiple donors to examine the consequences of APOBEC3 protein function at natural levels of expression. We demonstrate that APOBEC3 proteins impart a profound deficiency to reverse transcription from the initial stages of cDNA synthesis, as well as excessive cytidine deamination (hypermutation) of the DNAs that are synthesized. Experiments using viruses from transfected cells and a novel method for mapping the 3' termini of cDNAs indicate that the inhibition of reverse transcription is not limited to a few specific sites, arguing that APOBEC3 proteins impede enzymatic processivity. Detailed analyses of mutation spectra in viral cDNA strongly imply that one particular APOBEC3 protein, APOBEC3G, provides the bulk of the antiviral phenotype in CD4(+) T cells, with the effects of APOBEC3F and APOBEC3D being less significant. Taken together, we conclude that the dual mechanisms of action of APOBEC3 proteins combine to deliver more effective restriction of HIV-1 than either function would by itself.

Original languageEnglish
Article numberN/A
Pages (from-to)1508-1517
Number of pages10
JournalJournal of Virology
Volume87
Issue number3
Early online date14 Nov 2012
DOIs
Publication statusPublished - Feb 2013

Keywords

  • IMMUNODEFICIENCY-VIRUS TYPE-1
  • SINGLE-STRANDED-DNA
  • VIF PROTEIN
  • CBF-BETA
  • ALU RETROTRANSPOSITION
  • ANTIRETROVIRAL FACTOR
  • RETROVIRAL INFECTION
  • ANTIVIRAL ACTIVITY
  • HUMAN MACROPHAGES
  • ENZYME APOBEC3G

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