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Published online April 16, 2007
doi:10.1083/jcb.200609050
The Journal of Cell Biology, Vol. 177, No. 2, 329-341
The Rockefeller University Press, 0021-9525 $30.00
© 2007 Deneka et al.
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Article

 In macrophages, HIV-1 assembles into an intracellular plasma membrane domain containing the tetraspanins CD81, CD9, and CD53

Magdalena Deneka1,2, Annegret Pelchen-Matthews1,2, Rahel Byland1,2, Ezequiel Ruiz-Mateos1,2, and Mark Marsh1,2

1 Cell Biology Unit, Medical Research Council Laboratory for Molecular Cell Biology, and 2 Department of Biochemistry and Molecular Biology, University College London, London WC1E 6BT, England, UK

Correspondence to Mark Marsh: m.marsh{at}ucl.ac.uk

In macrophages, HIV-1 has been shown to bud into intracellular structures that contain the late endosome marker CD63. We show that these organelles are not endosomes, but an internally sequestered plasma membrane domain. Using immunofluorescence microscopy and immunoelectron microscopy, we find that HIV-1 buds into a compartment that contains the tetraspanins CD81, CD9, and CD53. On uninfected macrophages, these proteins are seen at the cell surface and in intracellular vacuole-like structures with a complex content of vesicles and interconnected membranes that lack endosome markers, including CD63. Significantly, these structures are accessible to small tracers (horseradish peroxidase or ruthenium red) applied to cells at 4°C, indicating that they are connected to the cell surface. HIV assembles on, and accumulates within, these intracellular compartments. Furthermore, CD63 is recruited to the virus-containing structures and incorporated into virions. These results indicate that, in macrophages, HIV-1 exploits a previously undescribed intracellular plasma membrane domain to assemble infectious particles.

M. Deneka and A. Pelchen-Matthews contributed equally to this paper.

Abbreviations used in this paper: DC, dendritic cell; HIV, human immunodeficiency virus type 1; MDM, monocyte-derived macrophages; MHC-II, major histocompatibility complex class II; MIIC, MHC-II compartment; MVB, multivesicular body; PAG, protein A-gold; PBMC, peripheral blood mononuclear cell; PI(4,5)P2, phosphatidylinositol 4,5-bis-phosphate; RR, ruthenium red; VCC, intracellular virus-containing compartment.


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