Selection and characterization of mercury-independent activation mutants of the Tn501 transcriptional regulator, MerR.
MerR is the transcriptional regulator of the mercury-resistance (mer) operon of transposon Tn501, acting at the mer promoter as both an activator in the presence of mercuric salts and a repressor in their absence. This paper reports a method for selection of constitutive activator mutants, which activate transcription in the absence of HgII, and the characterization of these MerRAC proteins. At least two mutations in the MerR protein were found necessary for strong mercury-independent activation, and these mutations lie in the C-terminal two-thirds of the MerR protein near the HgII-binding cysteines. A triple mutation was shown to increase activation over the corresponding double mutations. All mutant proteins caused further activation in the presence of HgII. The data support a mechanism in which a conformational change of one or both MerR subunits in the homodimer drives a distortion of DNA bound to a helix-turn-helix structure in the N-terminal region. A mutation in this putative helix-turn-helix region severely reduced both the repressor and activator functions of MerR.
Parkhill
Julian
J
Lawley
Blair
B
Hobman
Jonathan L
JL
Brown
Nigel L
NL
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001665:Binding Sites; D003545:Cysteine / Q000235:genetics; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D005819:Genetic Markers; D019077:Helix-Turn-Helix Motifs / Q000235:genetics; D008628:Mercury / Q000494:pharmacology*; D009154:Mutation / Q000235:genetics*; D010641:Phenotype; D012097:Repressor Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D020218:Response Elements / Q000235:genetics; D013816:Thermodynamics; D015534:Trans-Activators / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D005819:Genetic Markers; C051124:MerR protein, Bacteria; D012097:Repressor Proteins; D015534:Trans-Activators; D008628:Mercury; D003545:Cysteine