Gag induces the coalescence of clustered lipid rafts and tetraspanin-enriched microdomains at HIV-1 assembly sites on the plasma membrane

Ian Hogue, Jonathan R. Grover, Ferri Soheilian, Kunio Nagashima, Akira Ono

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

The HIV-1 structural protein Gag associates with two types of plasma membrane microdomains, lipid rafts and tetraspanin-enriched microdomains (TEMs), both of which have been proposed to be platforms for HIV-1 assembly. However, a variety of studies have demonstrated that lipid rafts and TEMs are distinct microdomains in the absence of HIV-1 infection. To measure the impact of Gag on microdomain behaviors, we took advantage of two assays: an antibody-mediated copatching assay and a Förster resonance energy transfer (FRET) assay that measures the clustering of microdomain markers in live cells without antibody-mediated patching. We found that lipid rafts and TEMs copatched and clustered to a greater extent in the presence of membrane-bound Gag in both assays, suggesting that Gag induces the coalescence of lipid rafts and TEMs. Substitutions in membrane binding motifs of Gag revealed that, while Gag membrane binding is necessary to induce coalescence of lipid rafts and TEMs, either acylation of Gag or binding of phosphatidylinositol-(4,5)-bisphosphate is sufficient. Finally, a Gag derivative that is defective in inducing membrane curvature appeared less able to induce lipid raft and TEM coalescence. A higher-resolution analysis of assembly sites by correlative fluorescence and scanning electron microscopy showed that coalescence of clustered lipid rafts and TEMs occurs predominately at completed cell surface virus-like particles, whereas a transmembrane raft marker protein appeared to associate with punctate Gag fluorescence even in the absence of cell surface particles. Together, these results suggest that different membrane microdomain components are recruited in a stepwise manner during assembly.

Original languageEnglish (US)
Pages (from-to)9749-9766
Number of pages18
JournalJournal of Virology
Volume85
Issue number19
DOIs
StatePublished - Oct 1 2011
Externally publishedYes

Fingerprint

Human immunodeficiency virus 1
HIV-1
plasma membrane
Cell Membrane
Lipids
lipids
Membrane Microdomains
Membranes
assays
Fluorescence
gag Gene Products
fluorescence
Acylation
Antibodies
Energy Transfer
acylation
Membrane Lipids
antibodies
Phosphatidylinositols
virus-like particles

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Gag induces the coalescence of clustered lipid rafts and tetraspanin-enriched microdomains at HIV-1 assembly sites on the plasma membrane. / Hogue, Ian; Grover, Jonathan R.; Soheilian, Ferri; Nagashima, Kunio; Ono, Akira.

In: Journal of Virology, Vol. 85, No. 19, 01.10.2011, p. 9749-9766.

Research output: Contribution to journalArticle

Hogue, Ian ; Grover, Jonathan R. ; Soheilian, Ferri ; Nagashima, Kunio ; Ono, Akira. / Gag induces the coalescence of clustered lipid rafts and tetraspanin-enriched microdomains at HIV-1 assembly sites on the plasma membrane. In: Journal of Virology. 2011 ; Vol. 85, No. 19. pp. 9749-9766.
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