Isolation and characterization of an oligomerization-negative mutant of HIV-1 integrase

Ganjam V. Kalpana, Alise Reicin, Grace S W Cheng, Mariya Sorin, Sandra Paik, Stephen P. Goff

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The yeast two-hybrid method was used to screen mutagenized DNAs to isolate a variant of the human immunodeficiency virus type 1 integrase (IN) that does not interact with the wild-type IN. The responsible mutation, leading to a single amino acid change (V260E) in the C-terminal domain of IN, blocks IN-IN multimerization but has only small effect on binding to a host interacting protein, INI1 (hSNF5). Binding studies in vitro confirmed the defect in multimerization of the mutant IN. Biochemical analyses of the mutant IN enzyme expressed in bacteria detected only subtle changes in its properties, suggesting that the yeast system is a sensitive reporter of correct IN conformation. Mutant virus carrying the V260E substitution was blocked in replication at the time of DNA integration, consistent with IN multimerization being important for its activity in vivo.

Original languageEnglish (US)
Pages (from-to)274-285
Number of pages12
JournalVirology
Volume259
Issue number2
DOIs
StatePublished - Jul 5 1999

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Integrases
Yeasts
p31 integrase protein, Human immunodeficiency virus 1
Two-Hybrid System Techniques
DNA
HIV-1
Viruses
Bacteria
Amino Acids
Mutation

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Isolation and characterization of an oligomerization-negative mutant of HIV-1 integrase. / Kalpana, Ganjam V.; Reicin, Alise; Cheng, Grace S W; Sorin, Mariya; Paik, Sandra; Goff, Stephen P.

In: Virology, Vol. 259, No. 2, 05.07.1999, p. 274-285.

Research output: Contribution to journalArticle

Kalpana, GV, Reicin, A, Cheng, GSW, Sorin, M, Paik, S & Goff, SP 1999, 'Isolation and characterization of an oligomerization-negative mutant of HIV-1 integrase', Virology, vol. 259, no. 2, pp. 274-285. https://doi.org/10.1006/viro.1999.9767
Kalpana, Ganjam V. ; Reicin, Alise ; Cheng, Grace S W ; Sorin, Mariya ; Paik, Sandra ; Goff, Stephen P. / Isolation and characterization of an oligomerization-negative mutant of HIV-1 integrase. In: Virology. 1999 ; Vol. 259, No. 2. pp. 274-285.
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