Summary
The nucleotide sequence of the putC region, from which divergent transcription of the putP and putA genes starts, was determined. The promoter region for the putA gene was restricted to the location between the HindIII site and the NcoI site or at the NcoI site by using putA-lacZ fusion plasmids and the transcriptional start for the putA gene was identified in the region between the HindIII site and the NcoI site by S1 mapping. This region also contains a potential CAP binding site, a ribosome binding site, and a sequence that is highly homologous to argTr. five potential promoters (putPp1-putPp5) for the putP gene, which were separate from the promoter region for the putA gene, were indicated by S1 mapping analysis of the putP gene transcripts. We concluded that the putC region is 419 bp long and contains two independent sets of promoters, regulating the expression of putP and putA genes in opposite directions. In addition, this region was found to contain an open reading frame (orf) capable of encoding a polypeptide of 111 amino acids in overlapping fashion. But studies using an orf-lacZ fusion gene showed that this open reading frame was not expressed.
Abbreviations
- CAP:
-
catabolite gene activator protein
- TTC:
-
2,3,5,-triphenyl tetrazolium chloride
References
Aiba H, Adhya S, de Crombrugghe B (1981) Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem 256:11905–11910
Ausubel FM (1984) Regulation of nitrogen fixation genes. Cell 37:5–6
Bochner BR, Savageau MA (1977) Generalized indicator plate for genetic, metabolic, and taxonomic studies with microorganisms. Appl Environ Microbiol 33:434–444
Casadaban MJ, Cohen SN (1980) Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol 138:179–207
Casadaban MJ, Chou J, Cohen SN (1980) In vitro gene fusions that join an enzymatically active β-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. j Bacteriol 143:971–980
Condamine H (1971) Mutans des voies de biosynthése et de dégradation de la proline chez E. coli K12. Ann Inst Pasteur (Paris) 120:9–22
Fowler AV, Zabin I (1977) The amino acid sequence of β-galactosidase of Escherichia coli. Proc Natl Acad Sci USA 74:1507–1510
Hawley DK, McClure WR (1983) Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res 11:2237–2255
Higgins CF, Ames GF-L (1982) Regulatory regions of two transport operons under nitrogen control: Nucleotide sequences. Proc Natl Acad Sci USA 79:1083–1087
Maloy S, Roth JR (1983) Regulation of proline utilization in Salmonella typhimurium: Characterization of put:: Mu d(Ap, lac) operon fusions. J Bacteriol 154:561–568
Menzel R, Roth J (1981) Regulation of the genes for proline utilization in Salmonella typhimurium: Autogenous repression by the putA gene product. J Mol Biol 148:21–44
Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101:20–79
Mogi T, Yamamoto H, Nakao T, Yamato I, Anraku Y (1986) Genetic and physical characterization of putP, the proline carrier gene of Escherichia coli K12. Mol Gen Genet 202:35–41
Nakao T, Yamato I, Anraku Y (1987) Nucleotide sequence of putP, the proline carrier gene of Escherichia coli K12. Mol Gen Genet 208:70–75
O'Neill MC, Amass K, de Crombrugghe B (1981) Molecular model of the DNA interaction site for the cyclic AMP receptor protein. Proc Natl Acad Sci USA 78:2213–2217
Ow DW, Ausubel FM (1983) Regulation of nitrogen metabolism genes by nifA gene product in Klebsiella pneumoniae. Nature 301:307–313
Prival MJ, Magasanik B (1971) Resistance to catabolite repression of histidase and proline oxidase during nitrogen-limited growth of Klebsiella aerogenes. J Biol Chem 246:6288–6296
Ratzkin B, Roth J (1978) Cluster of genes controlling proline degradation in Salmonella typhimurium. J Bacteriol 133:744–754
Ratzkin B, Grabnar M, Roth J (1978) Regulation of the major proline permease gene of Saomonella typhimurium. J Bacteriol 133:737–743
Sanger F, Nicklen S, Coulson A (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Shine J, Dalgarno L (1974) The 3′-terminal sequence of Escherichia coli 16S ribosomal RNA; complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346
Wood JM (1981) Genetics of L-proline utilization in Escherichia coli. J Bacteriol 146:895–901
Wood JM, Zadworny D (1979) Characterization of an inducible porter required for L-proline catabolism by Escherichia coli K12. Can J Biochem 57:1191–1199
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequence of the M13 mp18 and pUC19 vectors. Gene 33:103–119
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Communicated by M. Takanami
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Nakao, T., Yamato, I. & Anraku, Y. Nucleotide sequence of putC, the regulatory region for the put regulon of Escherichia coli K12. Molec Gen Genet 210, 364–368 (1987). https://doi.org/10.1007/BF00325707
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DOI: https://doi.org/10.1007/BF00325707