Synapsis in phage Bxb1 integration: selection mechanism for the correct pair of recombination sites

J Mol Biol. 2005 Jun 3;349(2):331-48. doi: 10.1016/j.jmb.2005.03.043. Epub 2005 Apr 7.

Abstract

Recombination by site-specific recombinases is a highly concerted process that requires synapsis of the correct pair of DNA substrates. Phage-encoded serine-integrases are unusual among the serine-recombinase family, which includes transposon resolvases and DNA invertases, in that they utilize two simple but different DNA substrates (attB and attP) and do not require accessory sites, additional proteins, or DNA supercoiling. Synapsis must therefore be directed solely by integrase-DNA interactions. We show here that the Bxb1 serine-integrase binds as a dimer to its two DNA substrates (attB, attP) and recombinant products (attL, attR) with similar affinities. However, synapsis occurs only between attP and attB, and not between any of the other nine possible site combinations. The Bxb1 integrase domain structure, the unusual DNA-binding properties of the integrase, and the characterization of a mutant protein with altered site-discrimination, are consistent with synaptic selectivity being derived from DNA sequence-induced changes in the conformations of integrase-DNA complexes.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Attachment Sites, Microbiological / genetics
  • Base Pairing / genetics
  • Base Sequence
  • DNA / genetics
  • DNA / metabolism
  • Dimerization
  • Integrases / metabolism
  • Molecular Sequence Data
  • Molecular Weight
  • Mutation / genetics
  • Mycobacteriophages / enzymology
  • Mycobacteriophages / genetics*
  • Protein Binding
  • Recombinases / chemistry
  • Recombinases / metabolism
  • Recombination, Genetic / genetics*
  • Sequence Alignment
  • Substrate Specificity
  • Tyrosine / genetics
  • Tyrosine / metabolism
  • Virus Integration / genetics*

Substances

  • Recombinases
  • Tyrosine
  • DNA
  • Integrases