Barrier-to-Autointegration Factor BAF Binds p55 Gag and Matrix and Is a Host Component of Human Immunodeficiency Virus Type 1 Virions

Malini Mansharamani, David R M Graham, Daphne Monie, Kenneth K. Lee, James Hildreth, Robert F. Siliciano, Katherine L. Wilson

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Abstract

Barrier-to-autointegration factor (BAF) is a conserved human chromatin protein exploited by retroviruses. Previous investigators showed that BAF binds double-stranded DNA nonspecifically and is a host component of preintegration complexes (PICs) isolated from cells infected with human immunodeficiency virus type 1 (HIV-1) or Moloney murine leukemia virus. BAF protects PIC structure and stimulates the integration of salt-stripped PICs into target DNA in vitro. PICs are thought to acquire BAF from the cytoplasm during infection. However, we identified two human tissues (of 16 tested) in which BAF mRNA was not detected: thymus and peripheral blood leukocytes, which are enriched in CD4+ T lymphocytes and macrophage precursors, respectively. BAF protein was detected in activated but not resting CD4+ T lymphocytes; thus, if BAF were essential for PIC function, we hypothesized that virions might "bring their own BAF." Supporting this model, BAF copurified with HIV-1 virions that were digested with subtilisin to remove microvesicle contaminants, and BAF was present in approximately zero to three copies per virion. In three independent assays, BAF bound directly to both p55 Gag (the structural precursor of HIV-1 virions) and its cleaved product, matrix. Using lysates from cells overexpressing Gag, endogenous BAF and Gag were coimmunoprecipitated by anti-bodies against Gag. Purified recombinant BAF had low micromolar affinities (1.1 to 1.4 μM) for recombinant Gag and matrix. We conclude that BAF is present at low levels in incoming virions, in addition to being acquired from the cytoplasm of newly infected cells. We further conclude that BAF might contribute to the assembly or activity of HIV-I PICs through direct binding to matrix, as well as DNA.

Original languageEnglish (US)
Pages (from-to)13084-13092
Number of pages9
JournalJournal of Virology
Volume77
Issue number24
DOIs
StatePublished - Dec 2003
Externally publishedYes

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ASJC Scopus subject areas

  • Immunology

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