LEDGF dominant interference proteins demonstrate prenuclear exposure of HIV-1 integrase and synergize with LEDGF depletion to destroy viral infectivity

Anne M. Meehan, Dyana T. Saenz, James Morrison, Chunling Hu, Mary Peretz, Eric M. Poeschla

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Target cell overexpression of the integrase binding domain (IBD) of LEDGF/p75 (LEDGF) inhibits HIV-1 replication. The mechanism and protein structure requirements for this dominant interference are unclear. More generally, how and when HIV-1 uncoating occurs postentry is poorly defined, and it is unknown whether integrase within the evolving viral core becomes accessible to cellular proteins prior to nuclear entry. We used LEDGF dominant interference to address the latter question while characterizing determinants of IBD antiviral activity. Fusions of green fluorescent protein (GFP) with multiple C-terminal segments of LEDGF inhibited HIV-1 replication substantially, but minimal chimeras of either polarity (GFP-IBD or IBD-GFP) were most effective. Combining GFP-IBD expression with LEDGF depletion was profoundly antiviral. CD4+ T cell lines were rendered virtually uninfectable, with single-cycle HIV-1 infectivity reduced 4 logs and high-input (multiplicity of infection = 5.0) replication completely blocked. We restricted GFP-IBD to specific intracellular locations and found that antiviral activity was preserved when the protein was confined to the cytoplasm or directed to the nuclear envelope. The life cycle block triggered by the cytoplasm-restricted protein manifested after nuclear entry, at the level of integration. We conclude that integrase within the viral core becomes accessible to host cell protein interaction in the cytoplasm. LEDGF dominant interference and depletion impair HIV-1 integration at distinct postentry stages. GFP-IBD may trigger premature or improper integrase oligomerization.

Original languageEnglish (US)
Pages (from-to)3570-3583
Number of pages14
JournalJournal of Virology
Volume85
Issue number7
DOIs
StatePublished - Apr 2011

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Integrases
Human immunodeficiency virus 1
crossover interference
green fluorescent protein
pathogenicity
Green Fluorescent Proteins
HIV-1
Proteins
cytoplasm
Protein Binding
proteins
Antiviral Agents
Cytoplasm
chimerism
nuclear membrane
protein structure
p31 integrase protein, Human immunodeficiency virus 1
lens epithelium-derived growth factor
life cycle (organisms)
T-lymphocytes

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

LEDGF dominant interference proteins demonstrate prenuclear exposure of HIV-1 integrase and synergize with LEDGF depletion to destroy viral infectivity. / Meehan, Anne M.; Saenz, Dyana T.; Morrison, James; Hu, Chunling; Peretz, Mary; Poeschla, Eric M.

In: Journal of Virology, Vol. 85, No. 7, 04.2011, p. 3570-3583.

Research output: Contribution to journalArticle

Meehan, Anne M. ; Saenz, Dyana T. ; Morrison, James ; Hu, Chunling ; Peretz, Mary ; Poeschla, Eric M. / LEDGF dominant interference proteins demonstrate prenuclear exposure of HIV-1 integrase and synergize with LEDGF depletion to destroy viral infectivity. In: Journal of Virology. 2011 ; Vol. 85, No. 7. pp. 3570-3583.
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