Abstract
The DNA sequence of the Salmonella typhimurium metA control region is presented. S1 nuclease mapping was used to determine the transcription initiation site. By measuring beta-galactosidase levels in Escherichia coli strains lysogenized with lambda phage carrying a metA-lacZ gene fusion, the MetR protein was shown to activate the metA gene. Homocysteine, an intermediate in methionine biosynthesis, plays a negative role in the MetR-mediated activation mechanism. Gel mobility shift assays and DNase I protection experiments showed that the MetR protein binds to a DNA fragment carrying the metA control region and protects a 26-bp region beginning 9 bp upstream of the -35 promoter sequence.
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Selected References
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- Baker R. F., Yanofsky C. The periodicity of RNA polymerase initiations: a new regulatory feature of transcription. Proc Natl Acad Sci U S A. 1968 May;60(1):313–320. doi: 10.1073/pnas.60.1.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
- Byerly K. A., Urbanowski M. L., Stauffer G. V. The metR binding site in the Salmonella typhimurium metH gene: DNA sequence constraints on activation. J Bacteriol. 1991 Jun;173(11):3547–3553. doi: 10.1128/jb.173.11.3547-3553.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cai X. Y., Maxon M. E., Redfield B., Glass R., Brot N., Weissbach H. Methionine synthesis in Escherichia coli: effect of the MetR protein on metE and metH expression. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4407–4411. doi: 10.1073/pnas.86.12.4407. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cai X. Y., Redfield B., Maxon M., Weissbach H., Brot N. The effect of homocysteine on MetR regulation of metE, metR and metH expression in vitro. Biochem Biophys Res Commun. 1989 Aug 30;163(1):79–83. doi: 10.1016/0006-291x(89)92101-3. [DOI] [PubMed] [Google Scholar]
- Casadaban M. J., Chou J., Cohen S. N. In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. J Bacteriol. 1980 Aug;143(2):971–980. doi: 10.1128/jb.143.2.971-980.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dev I. K., Harvey R. J. Regulation of synthesis of serine hydroxymethyltransferase in chemostat cultures of Escherichia coli. J Biol Chem. 1984 Jul 10;259(13):8394–8401. [PubMed] [Google Scholar]
- Dev I. K., Harvey R. J. Role of methionine in the regulation of the synthesis of serine hydroxymethyltransferase in Escherichia coli. J Biol Chem. 1984 Jul 10;259(13):8402–8406. [PubMed] [Google Scholar]
- Feiss M., Siegele D. A., Rudolph C. F., Frackman S. Cosmid DNA packaging in vivo. Gene. 1982 Feb;17(2):123–130. doi: 10.1016/0378-1119(82)90064-6. [DOI] [PubMed] [Google Scholar]
- Fried M., Crothers D. M. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Dec 11;9(23):6505–6525. doi: 10.1093/nar/9.23.6505. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garner M. M., Revzin A. A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system. Nucleic Acids Res. 1981 Jul 10;9(13):3047–3060. doi: 10.1093/nar/9.13.3047. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Greene R. C. Methionine limitation in Escherichia coli K-12 by growth on the sulfoxides of D-methionine. J Bacteriol. 1973 Oct;116(1):230–234. doi: 10.1128/jb.116.1.230-234.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
- Michaeli S., Mevarech M., Ron E. Z. Regulatory region of the metA gene of Escherichia coli K-12. J Bacteriol. 1984 Dec;160(3):1158–1162. doi: 10.1128/jb.160.3.1158-1162.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Panasenko S. M., Cameron J. R., Davis R. W., Lehman I. R. Five hundredfold overproduction of DNA ligase after induction of a hybrid lambda lysogen constructed in vitro. Science. 1977 Apr 8;196(4286):188–189. doi: 10.1126/science.322281. [DOI] [PubMed] [Google Scholar]
- Plamann M. D., Stauffer G. V. Regulation of the Escherichia coli glyA gene by the metR gene product and homocysteine. J Bacteriol. 1989 Sep;171(9):4958–4962. doi: 10.1128/jb.171.9.4958-4962.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
- Saint-Girons I., Parsot C., Zakin M. M., Bârzu O., Cohen G. N. Methionine biosynthesis in Enterobacteriaceae: biochemical, regulatory, and evolutionary aspects. CRC Crit Rev Biochem. 1988;23 (Suppl 1):S1–42. doi: 10.3109/10409238809083374. [DOI] [PubMed] [Google Scholar]
- Savin M. A., Flavin M., Slaughter C. Regulation of homocysteine biosynthesis in Salmonella typhimurium. J Bacteriol. 1972 Aug;111(2):547–556. doi: 10.1128/jb.111.2.547-556.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmitz A., Galas D. J. The interaction of RNA polymerase and lac repressor with the lac control region. Nucleic Acids Res. 1979 Jan;6(1):111–137. doi: 10.1093/nar/6.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shimada K., Weisberg R. A., Gottesman M. E. Prophage lambda at unusual chromosomal locations. I. Location of the secondary attachment sites and the properties of the lysogens. J Mol Biol. 1972 Feb 14;63(3):483–503. doi: 10.1016/0022-2836(72)90443-3. [DOI] [PubMed] [Google Scholar]
- Stauffer G. V., Plamann M. D., Stauffer L. T. Construction and expression of hybrid plasmids containing the Escherichia coli glyA genes. Gene. 1981 Jun-Jul;14(1-2):63–72. doi: 10.1016/0378-1119(81)90148-7. [DOI] [PubMed] [Google Scholar]
- Urbanowski M. L., Stauffer G. V. Autoregulation by tandem promoters of the Salmonella typhimurium LT2 metJ gene. J Bacteriol. 1986 Mar;165(3):740–745. doi: 10.1128/jb.165.3.740-745.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Urbanowski M. L., Stauffer G. V. Genetic and biochemical analysis of the MetR activator-binding site in the metE metR control region of Salmonella typhimurium. J Bacteriol. 1989 Oct;171(10):5620–5629. doi: 10.1128/jb.171.10.5620-5629.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Urbanowski M. L., Stauffer G. V. Role of homocysteine in metR-mediated activation of the metE and metH genes in Salmonella typhimurium and Escherichia coli. J Bacteriol. 1989 Jun;171(6):3277–3281. doi: 10.1128/jb.171.6.3277-3281.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Urbanowski M. L., Stauffer G. V. The control region of the metH gene of Salmonella typhimurium LT2: an atypical met promoter. Gene. 1988 Dec 15;73(1):193–200. doi: 10.1016/0378-1119(88)90325-3. [DOI] [PubMed] [Google Scholar]
- Urbanowski M. L., Stauffer L. T., Plamann L. S., Stauffer G. V. A new methionine locus, metR, that encodes a trans-acting protein required for activation of metE and metH in Escherichia coli and Salmonella typhimurium. J Bacteriol. 1987 Apr;169(4):1391–1397. doi: 10.1128/jb.169.4.1391-1397.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]