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Structure and function of the Pseudomonas putida integration host factor

R Calb1, A Davidovitch, S Koby

  • 1Department of Molecular Genetics and Biotechnology, Hadassah Medical School, The Hebrew University, Jerusalem, Israel.

Journal of Bacteriology
|November 1, 1996
PubMed
Summary
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Integration host factor (IHF) is crucial for toluene degradation in Pseudomonas putida. This study cloned P. putida IHF genes, showing they complement E. coli mutants and are essential for xyl operon expression.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Integration host factor (IHF) is a DNA-binding protein found in gram-negative bacteria.
  • IHF plays a role in DNA manipulation and gene regulation.

Purpose of the Study:

  • To clone, sequence, and functionally analyze the genes for the two subunits of IHF from Pseudomonas putida.
  • To investigate the role of IHF in the regulation of the xyl operon and toluene degradation.

Main Methods:

  • Gene cloning and sequencing of P. putida ihfA and ihfB.
  • Complementation analysis using E. coli ihf mutants.
  • Construction of a P. putida ihfA gene knockout mutant.
  • Analysis of xyl operon promoter activity and benzyl alcohol degradation.

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Main Results:

  • P. putida IHF subunits are similar in length to E. coli IHF subunits and can form functional heterodimers with E. coli IHF.
  • Amino acid differences are concentrated in specific domains, not affecting DNA binding or dimer formation.
  • IHF is essential for the expression of the P(U) promoter of the xyl operon.
  • An ihfA knockout mutant is defective in benzyl alcohol degradation, highlighting IHF's role in toluene degradation.

Conclusions:

  • The P. putida IHF protein is functionally conserved and plays a vital role in regulating the xyl operon.
  • IHF is essential for the degradation of aromatic compounds via the toluene degradation pathway in P. putida.