HIV-1 Protease Dimer Interface Mutations that Compensate for Viral Reverse Transcriptase Instability in Infectious Virions

Isabel Olivares, Alok Mulky, Peter I. Boross, J. Tőzsér, John C. Kappes, Cecilio López-Galíndez, Luis Menéndez-Arias

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Mature enzymes encoded within the human immunodeficiency virus type 1 (HIV-1) genome (protease (PR), reverse transcriptase (RT) and integrase (IN)) derive from proteolytic processing of a large polyprotein (Gag-Pol). Gag-Pol processing is catalyzed by the viral PR, which is active as a homodimer. The HIV-1 RT functions as a heterodimer (p66/p51) composed of subunits of 560 and 440 amino acid residues, respectively. Both subunits have identical amino acid sequence, but p51 lacks 120 residues that are removed by the HIV-1 PR during viral maturation. While p66 is the catalytic subunit, p51 has a primarily structural role. Amino acid substitutions affecting the stability of p66/p51 (i.e. F130W) have a deleterious effect on viral fitness. Previously, we showed that the effects of F130W are mediated by p51 and can be compensated by mutation T58S. While studying the dynamics of emergence of the compensatory mutation, we observed that mutations in the viral PR-coding region were selected in HIV clones containing the RT substitution F130W, before the imposition of T58S/F130W mutations. The PR mutations identified (G94S and T96S) improved the replication capacity of the F130W mutant virus. By using a trans-complementation assay, we demonstrate that the loss of p66/p51 heterodimer stability caused by Trp130 can be attributed to an increased susceptibility of RT to viral PR degradation. Recombinant HIV-1 PRs bearing mutations G94S or T96S showed decreased dimer stability and reduced catalytic efficiency. These results were consistent with crystallographic data showing the location of both residues in the PR dimerization interface.

Original languageEnglish
Pages (from-to)369-381
Number of pages13
JournalJournal of Molecular Biology
Volume372
Issue number2
DOIs
Publication statusPublished - Sep 14 2007

Fingerprint

RNA-Directed DNA Polymerase
Virion
HIV-1
Peptide Hydrolases
Mutation
pol Gene Products
gag Gene Products
Integrases
Dimerization
Amino Acid Substitution
Amino Acid Sequence
Catalytic Domain
Clone Cells
HIV
Genome
Viruses
Amino Acids
Enzymes

Keywords

  • fitness
  • HIV
  • protease
  • retrovirus
  • reverse transcriptase

ASJC Scopus subject areas

  • Virology

Cite this

HIV-1 Protease Dimer Interface Mutations that Compensate for Viral Reverse Transcriptase Instability in Infectious Virions. / Olivares, Isabel; Mulky, Alok; Boross, Peter I.; Tőzsér, J.; Kappes, John C.; López-Galíndez, Cecilio; Menéndez-Arias, Luis.

In: Journal of Molecular Biology, Vol. 372, No. 2, 14.09.2007, p. 369-381.

Research output: Contribution to journalArticle

Olivares, Isabel ; Mulky, Alok ; Boross, Peter I. ; Tőzsér, J. ; Kappes, John C. ; López-Galíndez, Cecilio ; Menéndez-Arias, Luis. / HIV-1 Protease Dimer Interface Mutations that Compensate for Viral Reverse Transcriptase Instability in Infectious Virions. In: Journal of Molecular Biology. 2007 ; Vol. 372, No. 2. pp. 369-381.
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