PMID:1452024
| Citation |
Loudon, JA and Loughlin, RE (1992) Mutagenesis and regulation of the cysJ promoter of Escherichia coli K-12. Gene 122:17-25 |
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| Abstract |
The cysJ promoter of Escherichia coli K-12, which is positively controlled by the CysB regulatory protein, was localized through the formation of a fusion of cysJ, the gene encoding NADPH-cytochrome c reductase with lacZ. The position of the transcription start point was determined and the orientation of transcription was shown to be counterclockwise on the E. coli K-12 map. Oligodeoxyribonucleotide-directed mutagenesis of the inferred -10 and -35 regions indicated that both sites could be altered to produce promoter 'down' mutations. When the -10 region was made to agree with the -10 consensus sequence, there was increased function under conditions of repression (that is, in the presence of cysteine). Upstream deletions, as well as mutations in a region proposed to be involved in binding of the CysB regulatory protein, identified sequences important for promoter activity from -90 to -78 and from -71 to -66. By comparison of the sequences of four cys promoters, a possible CysB-binding site was found which included the region shown to be required for the positive regulation of the cysJ promoter. |
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| Keywords |
Amino Acid Sequence; Bacterial Proteins/metabolism; Base Sequence; Binding Sites; DNA, Bacterial; DNA-Binding Proteins/metabolism; Escherichia coli/genetics; Gene Expression Regulation, Bacterial; Molecular Sequence Data; Mutagenesis, Site-Directed; Oxidoreductases Acting on Sulfur Group Donors/genetics; Plasmids; Promoter Regions, Genetic; Sequence Homology, Nucleic Acid; Sulfite Reductase (NADPH) |
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