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PMID: 17267500 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, N.I.H., Intramural Research Support, Non-U.S. Gov't

Downregulation of CD4 by human immunodeficiency virus type 1 Nef is dependent on clathrin and involves direct interaction of Nef with the AP2 clathrin adaptor.

Journal of virology ·Vol. 81 ·No. 8 ·2007-04-00 ·Pages 3877-90

Chaudhuri R, Lindwasser OW, Smith WJ, Hurley JH, Bonifacino JS

Abstract

Nef, an accessory protein of human and simian immunodeficiency viruses, is a critical determinant of pathogenesis that promotes the progression from infection to AIDS. The pathogenic effects of Nef are in large part dependent on its ability to downregulate the macrophage and T-cell coreceptor, CD4. It has been proposed that Nef induces downregulation by linking the cytosolic tail of CD4 to components of the host-cell protein trafficking machinery. To identify these components, we developed a novel Nef-CD4 downregulation system in Drosophila melanogaster S2 cells. We found that human immunodeficiency virus type 1 (HIV-1) Nef downregulates human CD4 in S2 cells and that this process is subject to the same sequence requirements as in human cells. An RNA interference screen targeting protein trafficking genes in S2 cells revealed a requirement for clathrin and the clathrin-associated, plasma membrane-localized AP2 complex in the downregulation of CD4. The requirement for AP2 was confirmed in the human cell line HeLa. We also used a yeast three-hybrid system and glutathione S-transferase pull-down analyses to demonstrate a robust, direct interaction between HIV-1 Nef and AP2. This interaction requires a dileucine motif in Nef that is also essential for downregulation of CD4. Together, these results support a model in which HIV-1 Nef downregulates CD4 by promoting its accelerated endocytosis by a clathrin/AP2 pathway.

MeSH Terms
Adaptor Protein Complex 2/metabolism Animals CD4 Antigens/biosynthesis Cell Line Clathrin/metabolism Down-Regulation Drosophila melanogaster Flow Cytometry Gene Products, nef/metabolism Gene Silencing HIV-1/physiology Humans Microscopy, Fluorescence Protein Binding Protein Interaction Mapping RNA Interference Transfection nef Gene Products, Human Immunodeficiency Virus
Chemicals
Adaptor Protein Complex 2 CD4 Antigens Clathrin Gene Products, nef nef Gene Products, Human Immunodeficiency Virus nef protein, Human immunodeficiency virus 1
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Chaudhuri Rittik
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Bldg. 18T, Rm. 101, National Institutes of Health, Bethesda, MD 20892, USA.
Lindwasser O Wolf
Smith William J
Hurley James H
Bonifacino Juan S
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.02725-06
Published
2007-04-00
Epub
2007-00-31
Pages
3877-90
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC1866153
Subset
IM
Grants
NICHD NIH HHS · K22 HD54602 · United States
Intramural NIH HHS · United States
External Links
PubMed source DOI: 10.1128/JVI.02725-06 PMC Full Text View journal details
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