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PMID: 20532212 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

The SOCS-box of HIV-1 Vif interacts with ElonginBC by induced-folding to recruit its Cul5-containing ubiquitin ligase complex.

PLoS pathogens ·Vol. 6 ·No. 6 ·2010-06-03 ·Pages e1000925

Bergeron JR, Huthoff H, Veselkov DA, Beavil RL, Simpson PJ, Matthews SJ, Malim MH, Sanderson MR

Abstract

The HIV-1 viral infectivity factor (Vif) protein recruits an E3 ubiquitin ligase complex, comprising the cellular proteins elongin B and C (EloBC), cullin 5 (Cul5) and RING-box 2 (Rbx2), to the anti-viral proteins APOBEC3G (A3G) and APOBEC3F (A3F) and induces their polyubiquitination and proteasomal degradation. In this study, we used purified proteins and direct in vitro binding assays, isothermal titration calorimetry and NMR spectroscopy to describe the molecular mechanism for assembly of the Vif-EloBC ternary complex. We demonstrate that Vif binds to EloBC in two locations, and that both interactions induce structural changes in the SOCS box of Vif as well as EloBC. In particular, in addition to the previously established binding of Vif's BC box to EloC, we report a novel interaction between the conserved Pro-Pro-Leu-Pro motif of Vif and the C-terminal domain of EloB. Using cell-based assays, we further show that this interaction is necessary for the formation of a functional ligase complex, thus establishing a role of this motif. We conclude that HIV-1 Vif engages EloBC via an induced-folding mechanism that does not require additional co-factors, and speculate that these features distinguish Vif from other EloBC specificity factors such as cellular SOCS proteins, and may enhance the prospects of obtaining therapeutic inhibitors of Vif function.

MeSH Terms
Amino Acid Sequence Cullin Proteins/chemistry,metabolism Elongin HIV Infections/metabolism,pathology HIV-1/metabolism Humans Immunoprecipitation Magnetic Resonance Spectroscopy Molecular Sequence Data Protein Folding Suppressor of Cytokine Signaling Proteins/chemistry,metabolism Transcription Factors/chemistry,metabolism Ubiquitin-Protein Ligases/metabolism Ubiquitination vif Gene Products, Human Immunodeficiency Virus/chemistry,metabolism
Chemicals
CUL5 protein, human Cullin Proteins ELOB protein, human ELOC protein, human Elongin Suppressor of Cytokine Signaling Proteins Transcription Factors vif Gene Products, Human Immunodeficiency Virus Ubiquitin-Protein Ligases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Bergeron Julien R C
Department of Infectious Diseases, King's College London School of Medicine, London, United Kingdom.
Huthoff Hendrik
Veselkov Dennis A
Beavil Rebecca L
Simpson Peter J
Matthews Stephen J
Malim Michael H
Sanderson Mark R
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Article Info
Journal
PLoS pathogens
Abbr.
PLoS Pathog
ISSN
1553-7374
DOI
10.1371/journal.ppat.1000925
Published
2010-06-03
Epub
2010-00-03
Pages
e1000925
Language
English
Region
United States
NLM ID
101238921
PMCID
PMC2880568
Subset
IM
Grants
Wellcome Trust · United Kingdom
Medical Research Council · G0401570 · United Kingdom
Wellcome Trust · 084280/Z/07/Z · United Kingdom
External Links
PubMed source DOI: 10.1371/journal.ppat.1000925 PMC Full Text View journal details
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