Two-step site selection for serine-integrase-mediated excision: DNA-directed integrase conformation and central dinucleotide proofreading

Proc Natl Acad Sci U S A. 2008 Mar 4;105(9):3238-43. doi: 10.1073/pnas.0711649105. Epub 2008 Feb 25.

Abstract

Bacteriophage-encoded serine-integrases are members of the large family of serine-recombinases and catalyze site-specific integrative recombination between a phage attP site and a bacterial attB site to form an integrated prophage. Prophage excision involves a second site-specific recombination event, in which the sites generated by integration, attL and attR, are used as substrates to regenerate attP and attB. Excision is catalyzed by integrase but also requires a phage-encoded recombination directionality factor (RDF). The Bxb1 recombination sites, attP and attB, are small (<50 bp), different in sequence, and quasisymmetrical, and they give rise to attL- and attR-recombinant products that are asymmetric but similar to each other, each being composed of B- and P-type half-sites. We show here that the determination of correct excision products is a two-step process, with a presynaptic RDF-dependent step that aligns attL and attR in the correct orientation and a postsynaptic step in which the nonpalindromic central dinucleotide confers identity to attL and attR and prevents each from recombining with itself.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Attachment Sites, Microbiological*
  • DNA
  • Integrases / chemistry
  • Integrases / metabolism*
  • Mycobacteriophages / enzymology
  • Nucleotides*
  • Protein Conformation
  • Recombination, Genetic*

Substances

  • Nucleotides
  • DNA
  • Integrases