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PMID: 20728451 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Molecular insight into the conformational dynamics of the Elongin BC complex and its interaction with HIV-1 Vif.

Journal of molecular biology ·Vol. 402 ·No. 5 ·2010-10-08 ·Pages 892-904

Marcsisin SR, Engen JR

Abstract

The human immunodeficiency virus type 1 virion infectivity factor (Vif) inhibits the innate viral immunity afforded by the APOBEC3 family of cytidine deaminases. Vif targets the APOBEC3 family for poly-ubiquitination and subsequent proteasomal degradation by linking the Elongin-BC-dependent ubiquitin ligase complex with the APOBEC3 proteins. The interaction between Vif and the heterodimeric Elongin BC complex, which is mediated by Vif's viral suppressor of cytokine signaling box, is essential for Vif function. The biophysical consequences of the full-length Vif:Elongin BC interaction have not been extensively reported. In this study, hydrogen exchange mass spectrometry was used to dissect the Vif:Elongin BC interaction. Elongin C was found to be highly dynamic in the Elongin BC complex while Elongin B was much more stable. Recombinant full-length Vif interacted with the Elongin BC complex in vitro with a K(d) of 1.9 μM and resulted in observable changes in deuterium uptake in both Elongin C and B. Upon binding to Elongin BC, no significant global conformational changes were detected in Vif by hydrogen exchange mass spectrometry, but a short fragment of Vif that consisted of the viral suppressor of cytokine signaling box showed decreased deuterium incorporation upon Elongin BC incubation, suggesting that this region folds upon binding.

MeSH Terms
Amino Acid Sequence Elongin Humans Hydrogen/metabolism Mass Spectrometry Models, Chemical Models, Molecular Molecular Sequence Data Protein Binding Protein Conformation Protein Folding Protein Interaction Mapping Protein Structure, Quaternary Transcription Factors/chemistry,metabolism vif Gene Products, Human Immunodeficiency Virus/chemistry,metabolism
Chemicals
ELOB protein, human ELOC protein, human Elongin Transcription Factors vif Gene Products, Human Immunodeficiency Virus vif protein, Human immunodeficiency virus 1 Hydrogen
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Marcsisin Sean R
Department of Chemistry and Chemical Biology and the Barnett Institute of Chemical and Biological Analysis, Northeastern University, Boston, MA 02115, USA.
Engen John R
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Article Info
Journal
Journal of molecular biology
Abbr.
J Mol Biol
ISSN
1089-8638
DOI
10.1016/j.jmb.2010.08.026
Published
2010-10-08
Epub
2010-00-20
Pages
892-904
Language
English
Region
England
NLM ID
2985088R
PMCID
PMC2949506
Subset
IM
Grants
NIGMS NIH HHS · R01 GM070590 · United States
NIGMS NIH HHS · R01 GM086507 · United States
NIGMS NIH HHS · R01-GM086507 · United States
NIGMS NIH HHS · R01-GM070590 · United States
External Links
PubMed source DOI: 10.1016/j.jmb.2010.08.026 PMC Full Text View journal details
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