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Protein-DNA complexes in mycobacteriophage L5 integrative recombination

C E Peña1, J M Kahlenberg, G F Hatfull

  • 1Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USA.

Journal of Bacteriology
|January 12, 1999
PubMed

Insights

Mycobacteriophage L5 integrase (Int-L5) and mycobacterial integration host factor (mIHF) form an intasome complex. This complex mediates site-specific DNA integration into Mycobacterium genomes by capturing the attB site.

Area of Science:

  • Molecular Biology
  • Genetics
  • Microbiology

Background:

  • Mycobacteriophage L5 integrates into Mycobacterium spp. genomes via site-specific recombination.
  • This process involves the phage L5 attP site, the mycobacterial attB site, phage integrase (Int-L5), and mycobacterial integration host factor (mIHF).

Purpose of the Study:

  • To elucidate the mechanism of intasome formation and its role in attB site recognition and integration.
  • To understand the specific roles of different attP binding sites in the assembly and function of the recombination complex.

Main Methods:

  • Biochemical assays to study the assembly of the intasome complex.
  • Analysis of protein-DNA interactions between attP, Int-L5, and mIHF.
  • Investigating the role of specific attP binding sites (P1-P2 and P4-P5) in recombination.

Main Results:

  • The phage attP site, Int-L5, and mIHF assemble into a functional intasome complex.
  • The intasome is capable of capturing the attB site and forming products of integration.
  • Specific binding sites within attP (P4-P5) are crucial for intasome core formation, while others (P1-P2) mediate attB interaction.

Conclusions:

  • The intasome serves as the active recombination complex for L5 phage integration.
  • Distinct DNA binding sites within attP dictate the formation of specific protein-DNA architectures essential for attB capture and integration.

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