title: A phage protein that binds φC31 integrase to switch its directionality.
abstract: The serine integrase, Int, from the Streptomyces phage φC31 mediates the integration and excision of the phage genome into and out of the host chromosome. Integrases usually require a recombination directionality factor (RDF) or Xis to control integration and excision and, as φC31 Int only mediates integration in the absence of other phage proteins, we sought to identify a φC31 RDF. Here we report that the φC31 early protein, gp3 activated attL x attR recombination and inhibited attP x attB recombination. Gp3 binds to Int in solution and when Int is bound to the attachment sites. Kinetic analysis of the excision reaction suggested that gp3 modifies the interactions between Int and the substrates to form an active recombinase. In the presence of gp3, Int assembles an excision synaptic complex and the accumulation of the integration complex is inhibited. The structure of the excision synaptic complex, like that of the hyperactive mutant of Int, IntE449K, appeared to be biased towards one that favours the production of correctly joined products. The functional properties of φC31 gp3 resemble those of the evolutionarily unrelated RDF from phage Bxb1, suggesting that these two RDFs have arisen through convergent evolution.
authors.affiliation: Institute of Medical Sciences, Foresterhill, University of Aberdeen, Aberdeen AB25 2ZD, UK.
authors.forename: Thanafez
authors.initials: T
authors.lastname: Khaleel
authors.forename: Ellen
authors.initials: E
authors.lastname: Younger
authors.forename: Andrew R
authors.initials: AR
authors.lastname: McEwan
authors.forename: Anpu S
authors.initials: AS
authors.lastname: Varghese
authors.forename: Margaret C M
authors.initials: MC
authors.lastname: Smith
mesh_terms: D001287:Attachment Sites, Microbiological; D004926:Escherichia coli / Q000821:virology; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D011995:Recombination, Genetic; D017108:Streptococcus Phages / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D016662:Virus Integration
chemical_list: D014764:Viral Proteins; D019426:Integrases