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Published online 30 May 2006. doi:10.1083/jcb.200508165
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 173, Number 5, 795-807
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Article

 Mapping of tetraspanin-enriched microdomains that can function as gateways for HIV-1

Sascha Nydegger3, Sandhya Khurana1,3, Dimitry N. Krementsov2,3, Michelangelo Foti4, and Markus Thali1,2,3

1 Graduate Program in Microbiology and Molecular Genetics and 2 Graduate Program in Cellular and Molecular Biology, 3 Department of Microbiology and Molecular Genetics, University of Vermont, Burlington, VT 05405
4 Department of Cellular Physiology and Metabolism, University of Geneva, CH-1211 Geneva, Switzerland

Correspondence to Markus Thali: markus.thali{at}uvm.edu

Specific spatial arrangements of proteins and lipids are central to the coordination of many biological processes. Tetraspanins have been proposed to laterally organize cellular membranes via specific associations with each other and with distinct integrins. Here, we reveal the presence of tetraspanin-enriched microdomains (TEMs) containing the tetraspanins CD9, CD63, CD81, and CD82 at the plasma membrane. Fluorescence and immunoelectron microscopic analyses document that the surface of HeLa cells is covered by several hundred TEMs, each extending over a few hundred nanometers and containing predominantly two or more tetraspanins. Further, we reveal that the human immunodeficiency virus type 1 (HIV-1) Gag protein, which directs viral assembly and release, accumulates at surface TEMs together with the HIV-1 envelope glycoprotein. TSG101 and VPS28, components of the mammalian ESCRT1 (endosomal sorting complex required for transport), which is part of the cellular extravesiculation machinery critical for HIV-1 budding, are also recruited to cell surface TEMs upon virus expression, suggesting that HIV-1 egress can be gated through these newly mapped microdomains.

S. Nydegger and S. Khurana contributed equally to this paper.

Abbreviations used in this paper: ESCRT, endosomal sorting complex required for transport; HIV-1, human immunodeficiency virus type 1; LE, late endosome; MVB, multivesicular body; TEM, tetraspanin-enriched microdomain; VLP, virus-like particle.


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