Naturally occurring amino acid polymorphisms in human immunodeficiency virus type 1 (HIV-1) Gag p7NC and the C-cleavage site impact Gag-Pol processing by HIV-1 protease

Maureen M. Goodenow, Gregory Bloom, Stephanie L. Rose, Steven M. Pomeroy, Patricia O. O'Brien, Elena E. Perez, John W. Sleasman, Ben M. Dunn

Research output: Contribution to journalArticle

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Abstract

Human immunodeficiency virus type 1 (HIV-1) protease activity is targeted at nine cleavage sites comprising different amino acid sequences in the viral Gag-Pol polyprotein. Amino acid polymorphisms in protease and in regions of Gag, particularly p7NC and the C-cleavage site between p2 and p7NC, occur in natural variants of HIV-1 within infected patients. Studies were designed to examine the role of natural polymorphisms in protease and to identify determinants in Gag that modulate protease processing activity. Closely related Gag-Pol regions from an HIV-1-infected mother and two children were evaluated for processing in an inducible expression system, for protease activity on cleavage-site analogues, and for impact on replication by recombinant viruses. Gag-Pol regions displayed one of three processing phenotypes based on the appearance of Gag intermediates and accumulation of mature p24CA. Gag-Pol regions that were processed rapidly to produce p24CA resulted in high-level replication by recombinant viruses, while slow-processing Gag-Pol variants resulted in recombinant viruses that replicated with reduced kinetics in both T cell lines and peripheral blood mononuclear cells. Direct impact by Gag sequences on processing by protease was assessed by construction of chimeric Gag-Pol regions and by site-directed mutagenesis. Optimal protease activity occurred when Gag and Pol regions were derived from the same gag-pol allele. Heterologous Gag regions generally diminished rates and extent of protease processing. Natural polymorphisms in novel positions in p7NC and the C-cleavage site have a dominant effect on protease processing activity. Accumulation of Gag products after processing at the C site appears to delay subsequent cleavage and production of mature p24CA.

Original languageEnglish
Pages (from-to)137-149
Number of pages13
JournalVirology
Volume292
Issue number1
DOIs
StatePublished - Sep 3 2002

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HIV-1
Peptide Hydrolases
Amino Acids
Virus Replication
pol Gene Products
gag Gene Products
Site-Directed Mutagenesis
Amino Acid Sequence
Blood Cells
Alleles
Mothers
Viruses
T-Lymphocytes
Phenotype
Cell Line

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Naturally occurring amino acid polymorphisms in human immunodeficiency virus type 1 (HIV-1) Gag p7NC and the C-cleavage site impact Gag-Pol processing by HIV-1 protease. / Goodenow, Maureen M.; Bloom, Gregory; Rose, Stephanie L.; Pomeroy, Steven M.; O'Brien, Patricia O.; Perez, Elena E.; Sleasman, John W.; Dunn, Ben M.

In: Virology, Vol. 292, No. 1, 03.09.2002, p. 137-149.

Research output: Contribution to journalArticle

Goodenow, Maureen M. ; Bloom, Gregory ; Rose, Stephanie L. ; Pomeroy, Steven M. ; O'Brien, Patricia O. ; Perez, Elena E. ; Sleasman, John W. ; Dunn, Ben M. / Naturally occurring amino acid polymorphisms in human immunodeficiency virus type 1 (HIV-1) Gag p7NC and the C-cleavage site impact Gag-Pol processing by HIV-1 protease. In: Virology. 2002 ; Vol. 292, No. 1. pp. 137-149.
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abstract = "Human immunodeficiency virus type 1 (HIV-1) protease activity is targeted at nine cleavage sites comprising different amino acid sequences in the viral Gag-Pol polyprotein. Amino acid polymorphisms in protease and in regions of Gag, particularly p7NC and the C-cleavage site between p2 and p7NC, occur in natural variants of HIV-1 within infected patients. Studies were designed to examine the role of natural polymorphisms in protease and to identify determinants in Gag that modulate protease processing activity. Closely related Gag-Pol regions from an HIV-1-infected mother and two children were evaluated for processing in an inducible expression system, for protease activity on cleavage-site analogues, and for impact on replication by recombinant viruses. Gag-Pol regions displayed one of three processing phenotypes based on the appearance of Gag intermediates and accumulation of mature p24CA. Gag-Pol regions that were processed rapidly to produce p24CA resulted in high-level replication by recombinant viruses, while slow-processing Gag-Pol variants resulted in recombinant viruses that replicated with reduced kinetics in both T cell lines and peripheral blood mononuclear cells. Direct impact by Gag sequences on processing by protease was assessed by construction of chimeric Gag-Pol regions and by site-directed mutagenesis. Optimal protease activity occurred when Gag and Pol regions were derived from the same gag-pol allele. Heterologous Gag regions generally diminished rates and extent of protease processing. Natural polymorphisms in novel positions in p7NC and the C-cleavage site have a dominant effect on protease processing activity. Accumulation of Gag products after processing at the C site appears to delay subsequent cleavage and production of mature p24CA.",
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