title: Genetic analysis of transcriptional activation and repression in the Tn21 mer operon.
abstract: Transcription of the Tn21 mercury resistance operon (mer) is controlled by the toxic metal cation Hg(II). This control is mediated by the product of the merR gene, a 144-amino-acid protein which represses transcription of the structural genes (merTPCAD) in the absence of Hg(II) and activates transcription in the presence of Hg(II). We have used a mer-lac transcriptional fusion to obtain regulatory mutants in this metal-responsive system. Some mutants were defective in Hg(II)-induced activation while retaining repression function (a- r+), others were defective in repression but not activation (a+ r-), and some had lost both functions (a- r-). Mutations in three of the four cysteine residues of merR resulted in complete loss of Hg(II)-inducible activation but retention of the repressor function, suggesting that these residues serve as ligands for Hg(II) in the activation process. Other lesions adjacent to or very near these cysteines exhibited severely reduced activation and also retained repressor function. There were two putative helix-turn-helix (HTH) domains in merR, and mutants in each had very different phenotypes. A partially dominant mutation in the more amino-terminal region of the two putative HTH regions resulted in loss of both activation and repression (a- r-), consistent with a role for this region in DNA binding. Mutations in the more centrally located HTH region resulted only in loss of Hg(II)-induced activation (a- r+). Lesions in the central and in the carboxy-terminal regions of merR exhibited both Hg(II)-independent and Hg(II)-dependent transcriptional activation, suggesting that elements important in the activation mechanism may be widely distributed in this relatively small protein. The sole cis-acting mutant obtained with this operon fusion strategy, a down-promoter mutation, lies in a highly conserved base in the -35 region of the merTPCAD promoter.
authors.affiliation: Department of Microbiology, University of Georgia, Athens 30602.
authors.forename: W
authors.initials: W
authors.lastname: Ross
authors.forename: S J
authors.initials: SJ
authors.lastname: Park
authors.forename: A O
authors.initials: AO
authors.lastname: Summers
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000145:classification / Q000235:genetics / Q000502:physiology; D004268:DNA-Binding Proteins / Q000145:classification / Q000235:genetics / Q000502:physiology; D004926:Escherichia coli / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D008628:Mercury / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation; D009876:Operon; D012097:Repressor Proteins / Q000145:classification / Q000235:genetics / Q000502:physiology; D013489:Suppression, Genetic; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D008628:Mercury