Analysis of HIV-1 gag protein interactions via biotin ligase tagging

Christopher Ritchie, Isabel Cylinder, Emily J. Platt, Eric Barklis

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

We have examined the interactions of wild-type (WT) and matrix protein-deleted (ΔMA) HIV-1 precursor Gag (PrGag) proteins in virus-producing cells using a biotin ligase-tagging approach. To do so, WT and ΔMA PrGag proteins were tagged with the Escherichia coli promiscuous biotin ligase (BirA*), expressed in cells, and examined. Localization patterns of PrGag proteins and biotinylated proteins overlapped, consistent with observations that BirA*-tagged proteins biotinylate neighbor proteins that are in close proximity. Results indicate that BirA*-tagged PrGag proteins biotinylated themselves as well as WT PrGag proteins in trans. Previous data have shown that the HIV-1 Envelope (Env) protein requires an interaction with MA for assembly into virions. Unexpectedly, ΔMA proteins biotinylated Env, whereas WT BirA*-tagged proteins did not, suggesting that the presence of MA made Env inaccessible to biotinylation. We also identified over 50 cellular proteins that were biotinylated by BirA*-tagged PrGag proteins. These included membrane proteins, cytoskeleton-associated proteins, nuclear transport factors, lipid metabolism regulators, translation factors, and RNA-processing proteins. The identification of these biotinylated proteins offers new insights into HIV-1 Gag protein trafficking and activities and provides new potential targets for antiviral interference.

Original languageEnglish (US)
Pages (from-to)3988-4001
Number of pages14
JournalJournal of Virology
Volume89
Issue number7
DOIs
StatePublished - 2015

Fingerprint

gag Gene Products
biotin
Ligases
Biotin
Human immunodeficiency virus 1
ligases
HIV-1
Proteins
proteins
Protein Transport
Biotinylation
Cell Nucleus Active Transport
Cytoskeleton
Lipid Metabolism
Virion
Antiviral Agents
Membrane Proteins
biotinylation
RNA
Escherichia coli

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Analysis of HIV-1 gag protein interactions via biotin ligase tagging. / Ritchie, Christopher; Cylinder, Isabel; Platt, Emily J.; Barklis, Eric.

In: Journal of Virology, Vol. 89, No. 7, 2015, p. 3988-4001.

Research output: Contribution to journalArticle

Ritchie, Christopher ; Cylinder, Isabel ; Platt, Emily J. ; Barklis, Eric. / Analysis of HIV-1 gag protein interactions via biotin ligase tagging. In: Journal of Virology. 2015 ; Vol. 89, No. 7. pp. 3988-4001.
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