DNA bending in the Sin recombination synapse: functional replacement of HU by IHF.
The serine recombinase Sin requires a non-specific DNA-bending protein such as Hbsu for activity at its recombination site resH. Hbsu, and Sin subunits bound at site II of resH, together regulate recombination, ensuring selectivity for directly repeated resH sites by specifying assembly of an intertwined synapse. To investigate the role of the DNA-bending protein in defining the architecture of the synapse, we constructed a chimaeric recombination site (resF) which allows Hbsu to be substituted by IHF, binding specifically between site I (the crossover site) and site II. Two Sin dimers and one IHF dimer can bind together to the closely adjoining sites in resF, forming folded complexes. The precise position of the IHF site within the site I-site II spacer determines the conformation of these complexes, and also the reactivity of the resF sites in recombination assays. The data suggest that a sharp bend with a specific geometry is required in the spacer DNA, to bring the Sin dimers at sites I and II together in the correct relative orientation for synapse assembly and regulation, consistent with our model for a highly condensed synapse in which Hbsu/IHF has a purely architectural function.
Rowland
Sally-J
SJ
University of Glasgow, Institute of Biomedical and Life Sciences, Division of Molecular Genetics, Anderson College, UK. sr2q@udcf.gla.ac.uk
Boocock
Martin R
MR
Stark
W Marshall
WM
D001287:Attachment Sites, Microbiological / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry*; D010582:Bacteriophage lambda / Q000235:genetics; D001665:Binding Sites; D004254:DNA Nucleotidyltransferases / Q000737:chemistry*; D004269:DNA, Bacterial / Q000737:chemistry*; D004278:DNA, Superhelical / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000737:chemistry*; D034982:Integration Host Factors / Q000737:chemistry*; D008958:Models, Molecular; D011487:Protein Conformation; D011995:Recombination, Genetic
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004278:DNA, Superhelical; D004268:DNA-Binding Proteins; D034982:Integration Host Factors; C040330:histone-like protein HU, bacteria; D004254:DNA Nucleotidyltransferases; C086893:sin protein, Staphylococcus aureus