The LexA repressor binds within the deep helical groove of the activated RecA filament.
The RecA protein of Escherichia coli, as a result of DNA damage, catalyzes the cleavage of its own repressor, the LexA protein, and thereby initiates the SOS response. Using a non-cleavable LexA mutant, we have obtained a co-complex of both the RecA and LexA proteins on DNA. Mass analysis using scanning transmission electron microscopy suggests that the site size of the LexA repressor on RecA is two, which would be consistent with a nearest-neighbor exclusion model for binding. Three-dimensional reconstruction of electron micrographs of these filaments shows that the LexA protein is bound in the deep groove of the RecA filament, with two strong regions of contact that span adjacent RecA protomers within the filament. One contact is consistent with a proposed LexA binding site in the RecA crystal structure. The other contact maps onto a region that has been postulated to be a second DNA-binding site within RecA, which can explain the inhibition of RecA cleavage of LexA by excess DNA.
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Department of Cell Biology and Neuroanatomy, University of Minnesota Medical School, Minneapolis 55455.
Egelman
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EH
D001426:Bacterial Proteins / Q000378:metabolism*; D001665:Binding Sites; D004249:DNA Damage; D004261:DNA Replication; D004269:DNA, Bacterial / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008433:Mathematics; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D008958:Models, Molecular; D011487:Protein Conformation; D011938:Rec A Recombinases / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D012097:Repressor Proteins / Q000378:metabolism*; D012697:Serine Endopeptidases; D000099160:LexA Repressor Protein
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D046911:Macromolecular Substances; D011938:Rec A Recombinases; D012097:Repressor Proteins; D012697:Serine Endopeptidases; D000099160:LexA Repressor Protein