A phage protein that binds [straight phi]C31 integrase to switch its directionality

• Published in: Molecular microbiology Vol. 80; no. 6; p. 1450
• Main Authors: Khaleel, Thanafez, Younger, Ellen, McEwan, Andrew R, Varghese, Anpu S, Smith, Margaret C M
• Format: Journal Article
• Language: English
• Published: Oxford Blackwell Publishing Ltd 01.06.2011
• Subjects: Binding sites; Evolution; Proteins
• ISSN: 0950-382X; 1365-2958

The serine integrase, Int, from the Streptomyces phage ...C31 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 ...C31 Int only mediates integration in the absence of other phage proteins, we sought to identify a ...C31 RDF. Here we report that the ...C31 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 ...C31 gp3 resemble those of the evolutionarily unrelated RDF from phage Bxb1, suggesting that these two RDFs have arisen through convergent evolution. (ProQuest: ... denotes formulae/symbols omitted.)