title: Identification of the CysB-regulated gene, hslJ, related to the Escherichia coli novobiocin resistance phenotype.
abstract: The cysB gene product is a LysR-type regulatory protein required for expression of the cys regulon. cysB mutants of Escherichia coli and Salmonella, along with being auxotrophs for the cysteine, exhibit increased resistance to the antibiotics novobiocin (Nov) and mecillinam. In this work, by using lambdaplacMu9 insertions creating random lacZ fusions, we identify a gene, hslJ, whose expression appeared to be increased in cysB mutants and needed for Nov resistance. Measurements of the HSLJ::lacZ gene fusion expression demonstrated that the hslJ gene is negatively regulated by CysB. In addition we observe the negative autogenous control of HslJ. When the control imposed by CysB is lifted in the cysB mutant, the elevation of Nov resistance can be achieved only in the presence of wild-type hslJ allele. A double cysB hslJ mutant restores the sensitivity to Nov. Overexpression of the wild-type HslJ protein either in a cysB(+) or a cysB(-) background increases the level of Nov resistance indicating that hslJ product is indeed involved in accomplishing this phenotype. The HSLJ::OmegaKan allele encodes the C-terminaly truncated mutant protein HslJ Q121Ter which is not functional in achieving the Nov resistance but when overexpressed induces the psp operon. Finally, we found that inactivation of hslJ does not affect the increased resistance to mecillinam in cysB mutants.
authors.affiliation: Institute of Molecular Genetics and Genetic Engineering, PO Box 446, 11001, Belgrade, Serbia and Montenegro, Yugoslavia.
authors.forename: Mirjana
authors.initials: M
authors.lastname: Lilic
authors.forename: Milija
authors.initials: M
authors.lastname: Jovanovic
authors.forename: Goran
authors.initials: G
authors.lastname: Jovanovic
authors.forename: Dragutin J
authors.initials: DJ
authors.lastname: Savic
mesh_terms: D000560:Amdinocillin / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D005809:Genes, Regulator; D018869:Heat-Shock Response / Q000235:genetics; D007763:Lac Operon; D009675:Novobiocin / Q000494:pharmacology*; D010406:Penicillins / Q000494:pharmacology; D010641:Phenotype; D011993:Recombinant Fusion Proteins / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; C062886:CysB protein, Bacteria; D010406:Penicillins; D011993:Recombinant Fusion Proteins; D009675:Novobiocin; D000560:Amdinocillin