Characterization of the mycobacteriophage L5 attachment site, attP

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...

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Bibliographic Details
Published in:Journal of molecular biology 1997-02, Vol.266 (1), p.76-92
Main Authors: Peña, C E, Lee, M H, Pedulla, M L, Hatfull, G F
Format: Article
Language:English
Subjects:
Bacteriophages - genetics
Bacteriophages - physiology
Base Sequence
Binding Sites
Chromosomes, Bacterial
Cloning, Molecular
Consensus Sequence
Deoxyribonuclease I
DNA Footprinting
DNA Mutational Analysis
Escherichia coli
Genome, Viral
Integrases - metabolism
Molecular Sequence Data
Mycobacterium - genetics
Mycobacterium - virology
Recombination, Genetic
Sequence Deletion
Virus Integration
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Summary: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.
ISSN:0022-2836
1089-8638
DOI:10.1006/jmbi.1996.0774