Contribution of E3-ubiquitin ligase activity to HIV-1 restriction by TRIM5α rh: Structure of the RING domain of TRIM5α

Maritza Lienlaf, Fumiaki Hayashi, Francesca Di Nunzio, Naoya Tochio, Takanori Kigawa, Shigeyuki Yokoyama, Felipe Diaz-Griffero

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

TRIM5α rh is a cytosolic protein that potently restricts HIV-1 before reverse transcription. TRIM5α rh is composed of four different domains: RING, B-box 2, coiled coil, and B30.2(SPRY). The contribution of each of these domains to restriction has been extensively studied, with the exception of the RING domain. The RING domain of TRIM5α exhibits E3-ubiquitin ligase activity, but the contribution of this activity to the restriction of HIV-1 is not known. To test the hypothesis that the E3-ubiquitin ligase activity of the RING domain modulates TRIM5α rh restriction of HIV-1, we correlated the E3-ubiquitin ligase activity of a panel of TRIM5α rh RING domain variants with the ability of these mutant proteins to restrict HIV-1. For this purpose, we first solved the nuclear magnetic resonance structure of the RING domain of TRIM5α and defined potential functional regions of the RING domain by homology to other RING domains. With this structural information, we performed a systematic mutagenesis of the RING domain regions and tested the TRIM5α RING domain variants for the ability to undergo self-ubiquitylation. Several residues, particularly the ones on the E2-binding region of the RING domain, were defective in their self-ubiquitylation ability. To correlate HIV-1 restriction to self-ubiquitylation, we used RING domain mutant proteins that were defective in self-ubiquitylation but preserve important properties required for potent restriction by TRIM5α rh, such as capsid binding and higher-order self-association. From these investigations, we found a set of residues that when mutated results in TRIM5α molecules that lost both the ability to potently restrict HIV-1 and their self-ubiquitylation activity. Remarkably, all of these changes were in residues located in the E2-binding region of the RING domain. Overall, these results demonstrate a role for TRIM5α self-ubiquitylation in the ability of TRIM5α to restrict HIV-1.

Original languageEnglish (US)
Pages (from-to)8725-8737
Number of pages13
JournalJournal of Virology
Volume85
Issue number17
DOIs
StatePublished - Sep 2011

Fingerprint

ubiquitin-protein ligase
Ubiquitin-Protein Ligases
Human immunodeficiency virus 1
Ubiquitination
HIV-1
Mutant Proteins
mutants
reverse transcription
capsid
proteins
Capsid
mutagenesis
Mutagenesis
Reverse Transcription
preserves
nuclear magnetic resonance spectroscopy
Magnetic Resonance Spectroscopy

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Contribution of E3-ubiquitin ligase activity to HIV-1 restriction by TRIM5α rh : Structure of the RING domain of TRIM5α. / Lienlaf, Maritza; Hayashi, Fumiaki; Di Nunzio, Francesca; Tochio, Naoya; Kigawa, Takanori; Yokoyama, Shigeyuki; Diaz-Griffero, Felipe.

In: Journal of Virology, Vol. 85, No. 17, 09.2011, p. 8725-8737.

Research output: Contribution to journalArticle

Lienlaf, Maritza ; Hayashi, Fumiaki ; Di Nunzio, Francesca ; Tochio, Naoya ; Kigawa, Takanori ; Yokoyama, Shigeyuki ; Diaz-Griffero, Felipe. / Contribution of E3-ubiquitin ligase activity to HIV-1 restriction by TRIM5α rh : Structure of the RING domain of TRIM5α. In: Journal of Virology. 2011 ; Vol. 85, No. 17. pp. 8725-8737.
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