Abstract
We isolated an Escherichia coli methionine auxotroph that displays a growth phenotype similar to that of known metF mutants but has elevated levels of 5,10-methylenetetrahydrofolate reductase, the metF gene product. Transduction analysis indicates that the mutant carries normal metE, metH, and metF genes; the phenotype is due to a single mutation, eliminating the possibility that the strain is a metE metH double mutant; and the new mutation is linked to the metE gene by P1 transduction. Plasmids carrying the Salmonella typhimurium metE gene and flanking regions complement the mutation, even when the plasmid-borne metE gene is inactivated. Enzyme assays show that the mutation results in a dramatic decrease in metE gene expression, a moderate decrease in metH gene expression, and a disruption of the metH-mediated vitamin B12 repression of the metE and metF genes. Our evidence suggests that the methionine auxotrophy caused by the new mutation is a result of insufficient production of both the vitamin B12-independent (metE) and vitamin B12-dependent (metH) transmethylase enzymes that are necessary for the synthesis of methionine from homocysteine. We propose that this mutation defines a positive regulatory gene, designated metR, whose product acts in trans to activate the metE and metH genes.
Full text
PDF






Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ahmed A. Mechanism of repression of methionine biosynthesis in Escherichia coli. I. The role of methionine, s-adenosylmethionine, and methionyl-transfer ribonucleic acid in repression. Mol Gen Genet. 1973 Jul 16;123(4):299–324. doi: 10.1007/BF00433648. [DOI] [PubMed] [Google Scholar]
- Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [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]
- Casadaban M. J. Fusion of the Escherichia coli lac genes to the ara promoter: a general technique using bacteriophage Mu-1 insertions. Proc Natl Acad Sci U S A. 1975 Mar;72(3):809–813. doi: 10.1073/pnas.72.3.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dawes J., Foster M. A. Vitamin B 12 and methionine synthesis in Escherichia coli. Biochim Biophys Acta. 1971 Jun 22;237(3):455–464. doi: 10.1016/0304-4165(71)90263-7. [DOI] [PubMed] [Google Scholar]
- Kung H. F., Spears C., Greene R. C., Weissbach H. Regulation of the terminal reactions in methionine biosynthesis by vitamin B 12 and methionine. Arch Biochem Biophys. 1972 May;150(1):23–31. doi: 10.1016/0003-9861(72)90005-7. [DOI] [PubMed] [Google Scholar]
- Kutzbach C., Stokstad E. L. Mammalian methylenetetrahydrofolate reductase. Partial purification, properties, and inhibition by S-adenosylmethionine. Biochim Biophys Acta. 1971 Dec 15;250(3):459–477. doi: 10.1016/0005-2744(71)90247-6. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Milner L., Whitfield C., Weissbach H. Effect of L-methionine and vitamin B 12 on methionine biosynthesis in Escherichia coli. Arch Biochem Biophys. 1969 Sep;133(2):413–419. doi: 10.1016/0003-9861(69)90470-6. [DOI] [PubMed] [Google Scholar]
- Mulligan J. T., Margolin W., Krueger J. H., Walker G. C. Mutations affecting regulation of methionine biosynthetic genes isolated by use of met-lac fusions. J Bacteriol. 1982 Aug;151(2):609–619. doi: 10.1128/jb.151.2.609-619.1982. [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]
- Raibaud O., Schwartz M. Positive control of transcription initiation in bacteria. Annu Rev Genet. 1984;18:173–206. doi: 10.1146/annurev.ge.18.120184.001133. [DOI] [PubMed] [Google Scholar]
- Schulte L. L., Stauffer L. T., Stauffer G. V. Cloning and characterization of the Salmonella typhimurium metE gene. J Bacteriol. 1984 Jun;158(3):928–933. doi: 10.1128/jb.158.3.928-933.1984. [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]
- Smith D. A., Childs J. D. Methionine genes and enzymes of Salmonella typhimurium. Heredity (Edinb) 1966 May;21(2):265–286. doi: 10.1038/hdy.1966.22. [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. The metH gene from Salmonella typhimurium LT2: cloning and initial characterization. Gene. 1986;44(2-3):211–217. doi: 10.1016/0378-1119(86)90184-8. [DOI] [PubMed] [Google Scholar]
- Whitehouse J. M., Smith D. A. Methionine and vitamin B 12 repression and precursor induction in the regulation of homocysteine methylation in Salmonella typhimurium. Mol Gen Genet. 1973;120(4):341–353. doi: 10.1007/BF00268148. [DOI] [PubMed] [Google Scholar]
- Whitfield C. D., Steers E. J., Jr, Weissbach H. Purification and properties of 5-methyltetrahydropteroyltriglutamate-homocysteine transmethylase. J Biol Chem. 1970 Jan 25;245(2):390–401. [PubMed] [Google Scholar]