Role of TRIM5α RING domain E3 ubiquitin ligase activity in capsid disassembly, reverse transcription blockade, and restriction of simian immunodeficiency virus

J Virol. 2011 Aug;85(16):8116-32. doi: 10.1128/JVI.00341-11. Epub 2011 Jun 15.

Abstract

The mammalian tripartite motif protein, TRIM5α, recognizes retroviral capsids entering the cytoplasm and blocks virus infection. Depending on the particular TRIM5α protein and retrovirus, complete disruption of the TRIM5α RING domain decreases virus-restricting activity to various degrees. TRIM5α exhibits RING domain-dependent E3 ubiquitin ligase activity, but the specific role of this activity in viral restriction is unknown. We created a panel of African green monkey TRIM5α (TRIM5α(AGM)) mutants, many of which are specifically altered in RING domain E3 ubiquitin ligase function, and characterized the phenotypes of these mutants with respect to restriction of simian and human immunodeficiency viruses (SIV(mac) and HIV-1, respectively). TRIM5α(AGM) ubiquitin ligase activity was essential for both the accelerated disassembly of SIV(mac) capsids and the disruption of reverse transcription. The levels of SIV(mac) particulate capsids in the cytosol of target cells expressing the TRIM5α variants strongly correlated with the levels of viral late reverse transcripts. RING-mediated ubiquitylation and B30.2(SPRY) domain-determined capsid binding independently contributed to the potency of SIV(mac) restriction by TRIM5α(AGM). In contrast, TRIM5α proteins attenuated in RING ubiquitin ligase function still accelerated HIV-1 capsid disassembly, inhibited reverse transcription, and blocked infection. Replacement of the helix-4/5 loop in the SIV(mac) capsid with the corresponding region of the HIV-1 capsid diminished the dependence of restriction on TRIM5α RING function. Thus, ubiquitylation mediated by the RING domain of TRIM5α(AGM) is essential for blocking SIV(mac) infection at the stage of capsid uncoating.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Restriction Factors
  • Capsid / metabolism
  • Capsid Proteins / genetics
  • Capsid Proteins / metabolism*
  • Carrier Proteins / immunology
  • Carrier Proteins / metabolism*
  • Chlorocebus aethiops
  • Dogs
  • HEK293 Cells
  • HIV-1 / genetics
  • HeLa Cells
  • Humans
  • Membrane Proteins / metabolism
  • RING Finger Domains
  • Reverse Transcription
  • Simian Immunodeficiency Virus / genetics
  • Simian Immunodeficiency Virus / metabolism*
  • Tripartite Motif Proteins
  • Ubiquitin-Protein Ligases / metabolism*
  • Ubiquitination

Substances

  • Antiviral Restriction Factors
  • Capsid Proteins
  • Carrier Proteins
  • Membrane Proteins
  • Tripartite Motif Proteins
  • TRIM5 protein, human
  • Ubiquitin-Protein Ligases