The LysR-type transcriptional regulator CysB controls the repression of hslJ transcription in Escherichia coli

• Published in: Microbiology (Society for General Microbiology) 2003-12, Vol.149 (12), p.3449-3459
• Main Authors: Jovanovic, Milija, Lilic, Mirjana, Savic, Dragutin J, Jovanovic, Goran
• Format: Article
• Language: English
• Subjects: Alleles; Amino Acid Sequence; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Base Sequence; Binding Sites - genetics; Biological and medical sciences; DNA, Bacterial - genetics; DNA, Bacterial - metabolism; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Bacterial; Genes, Bacterial; Genetic Complementation Test; Microbiology; Miscellaneous; Molecular Sequence Data; Mutation; Promoter Regions, Genetic; Transcription Factors - genetics; Transcription Factors - metabolism
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/mic.0.26609-0

Institute of Molecular Genetics and Genetic Engineering, Vojvode Stepe 444A, PO Box 446, 11001 Belgrade, Serbia and Montenegro Correspondence Goran Jovanovic mgjovano{at}eunet.yu The LysR-type transcriptional regulator (LTTR) CysB is a transcription factor in Escherichia coli cells, where as a homotetramer it binds the target promoter regions and activates the genes involved in sulphur utilization and sulphonate-sulphur metabolism, while negatively autoregulating its own transcription. The hslJ gene was found to be negatively regulated by CysB and directly correlated with novobiocin resistance of the bacterium. cysB mutants showed upregulation of the hslJ : : lacZ gene fusion and exhibited increased novobiocin resistance. In this study the hslJ transcription start point and the corresponding putative 70 promoter were determined. The hslJ promoter region was defined by employing different hslJ – lacZ operon fusions, and transcription of the hslJ gene was shown to be subject to both repression imposed by the CysB regulator and direct or indirect autogenous negative control. These two regulations compete to some extent but they are not mutually exclusive. CysB acts as a direct repressor of hslJ transcription and binds the hslJ promoter region that carries the putative CysB repressor site. This CysB binding, apparently responsible for repression, is enhanced in the presence of the ligand N -acetylserine (NAS), hitherto considered to be a positive cofactor in CysB-mediated gene regulations. Interallelic complementation of characterized CysB mutants I33N and S277Ter partially restored the repression of hslJ transcription and the consequent novobiocin sensitivity, but did not complement the cysteine auxotrophy. Abbreviations: HTH, helix–turn–helix; IR, inverted repeat; LTTR, LysR-type transcriptional regulator; NAS, N -acetylserine; OMP, outer-membrane protein; TSP, transcription start point; WT, wild-type Present address: Laboratory of Structural Microbiology, RU Box 52, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. Present address: Department of Microbiology and Immunology, Health Sciences Center, University of Oklahoma, PO Box 26901, Oklahoma City, OK 73190, USA.