Genetic analysis of homomeric interactions of human immunodeficiency virus type 1 integrase using the yeast two-hybrid system

Ganjam V. Kalpana, Stephen P. Goff

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

The retroviral integrase protein (IN) is responsible for catalyzing a concerted integration reaction in which the two termini of linear viral DNA are joined to host DNA. To probe the potential for IN to form protein multimers, we used the yeast two-hybrid system. The coexpression of a GAL4 DNA binding domain-IN fusion and a GAL4 activation domain-IN fusion together resulted in the successful activation of a GAL4-responsive LacZ reporter gene. The system was used to examine a variety of IN deletion mutants. The results suggest that the central region of the protein is necessary for multimerization and that the N-terminal zinc finger region is not important.

Original languageEnglish (US)
Pages (from-to)10593-10597
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number22
StatePublished - Nov 15 1993
Externally publishedYes

Fingerprint

Integrases
Two-Hybrid System Techniques
HIV-1
Proteins
Lac Operon
DNA
Zinc Fingers
Viral DNA
Reporter Genes

Keywords

  • GAL4
  • Integration
  • Protein-protein interactions
  • Retroviral replication

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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AB - The retroviral integrase protein (IN) is responsible for catalyzing a concerted integration reaction in which the two termini of linear viral DNA are joined to host DNA. To probe the potential for IN to form protein multimers, we used the yeast two-hybrid system. The coexpression of a GAL4 DNA binding domain-IN fusion and a GAL4 activation domain-IN fusion together resulted in the successful activation of a GAL4-responsive LacZ reporter gene. The system was used to examine a variety of IN deletion mutants. The results suggest that the central region of the protein is necessary for multimerization and that the N-terminal zinc finger region is not important.

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