lac operator DNA modification in the presence of proteolytic fragments of the repressor protein.
Singly end-labeled DNA fragments containing the lactose operator were methylated in the presence of the lactose repressor and homogeneous preparations of its proteolytic fragments. Binding of core protein produced by mild trypsin digestion yielded a methylation perturbation pattern that differed significantly from that elicited by binding to intact repressor, although similarities in the patterns for these related proteins were noted in the central, asymmetric region of the operator. An NH2-terminal peptide (residues 1 to 56) from lac repressor bound operator fragments in a nitrocellulose filter assay, but failed to perturb DNA methylation significantly relative to the pattern in the absence of peptide. Binding of hybrid tetramers of core and intact repressor monomers produced related but unique methylation patterns for the purines on the operator fragment. The general pattern of perturbation observed suggests preferred binding of a single NH2 terminus to the promoter-distal region of the operator and asymmetric interaction of the core region with the operator sequence. Differences in purine methylation patterns produced by the presence of effector complexes of repressor and core protein suggest the possible nature of changes in protein topology that result in the affinity changes accompanying induction.
D000477:Alkylating Agents; D001345:Autoradiography; D001483:Base Sequence; D004269:DNA, Bacterial / Q000378:metabolism*; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000032:analysis; D007763:Lac Operon; D008745:Methylation; D010446:Peptide Fragments / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism*; D013463:Sulfuric Acid Esters; D014157:Transcription Factors / Q000378:metabolism*; D014357:Trypsin
D000477:Alkylating Agents; D004269:DNA, Bacterial; D010446:Peptide Fragments; D012097:Repressor Proteins; D013463:Sulfuric Acid Esters; D014157:Transcription Factors; D014357:Trypsin; C007482:dimethyl sulfate