Characterization of the mycobacteriophage L5 attachment site, attP

J Mol Biol. 1997 Feb 14;266(1):76-92. doi: 10.1006/jmbi.1996.0774.

Abstract

Lysogenization of mycobacteriophage L5 involves integration of the phage genome into the Mycobacterium smegmatis chromosome. Integration occurs by a site-specific recombination event between a phage attachment site, attP, and a bacterial attachment site, attB, which is catalyzed by the phage-encoded integrase protein. DNase I footprinting reveals that L5 integrase binds to two types of sites within attP which span an unexpectedly large region of 413 bp: seven arm-type sites (P1 to P7) each of which correspond to a consensus sequence 5'-TGCaaCtcYy, and core-type sites at the points of strand exchange. Mutational analyses indicate that not all of the arm-type sites are required for integration, and that the P3 site and the rightmost pair of sites (P6 and P7) are dispensable for integration. We show that a 252 bp segment of attP DNA is sufficient for efficient integrative recombination and that int can be provided in trans for simple and efficient transformation of the mycobacteria.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacteriophages / genetics*
  • Bacteriophages / physiology
  • Base Sequence
  • Binding Sites
  • Chromosomes, Bacterial
  • Cloning, Molecular
  • Consensus Sequence
  • DNA Footprinting
  • DNA Mutational Analysis
  • Deoxyribonuclease I
  • Escherichia coli
  • Genome, Viral
  • Integrases / metabolism*
  • Molecular Sequence Data
  • Mycobacterium / genetics
  • Mycobacterium / virology*
  • Recombination, Genetic
  • Sequence Deletion
  • Virus Integration*

Substances

  • Integrases
  • Deoxyribonuclease I

Associated data

  • GENBANK/X70352
  • GENBANK/Z18946