Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Apr;174(7):2059–2064. doi: 10.1128/jb.174.7.2059-2064.1992

The mtrAB operon of Bacillus subtilis encodes GTP cyclohydrolase I (MtrA), an enzyme involved in folic acid biosynthesis, and MtrB, a regulator of tryptophan biosynthesis.

P Babitzke 1, P Gollnick 1, C Yanofsky 1
PMCID: PMC205820  PMID: 1551827

Abstract

mtrA of Bacillus subtilis was shown to be the structural gene for GTP cyclohydrolase I, an enzyme essential for folic acid biosynthesis. mtrA is the first gene in a bicistronic operon that includes mtrB, a gene involved in transcriptional attenuation control of the trp genes. mtrA of B. subtilis encodes a 20-kDa polypeptide that is 50% identical to rat GTP cyclohydrolase I. Increased GTP cyclohydrolase I activity was readily detected in crude extracts of B. subtilis and Escherichia coli in which MtrA was overproduced. Biochemical evidence indicating that MtrA catalyzes dihydroneopterin triphosphate and formic acid formation from guanosine triphosphate is presented. It was also shown that mtrB of B. subtilis encodes a 6-kDa polypeptide. Expression of mtrB is sufficient for transcriptional attenuation control of the B. subtilis trp gene cluster in Escherichia coli. Known interrelationships between genes involved in folic acid and aromatic amino acid biosynthesis in B. subtilis are described.

Full text

PDF
2059

Images in this article

2061Image
on p.2061

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anagnostopoulos C., Spizizen J. REQUIREMENTS FOR TRANSFORMATION IN BACILLUS SUBTILIS. J Bacteriol. 1961 May;81(5):741–746. doi: 10.1128/jb.81.5.741-746.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anthony C., Zatman L. J. The microbial oxidation of methanol. The prosthetic group of the alcohol dehydrogenase of Pseudomonas sp. M27: a new oxidoreductase prosthetic group. Biochem J. 1967 Sep;104(3):960–969. doi: 10.1042/bj1040960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burg A. W., Brown G. M. The biosynthesis of folic acid. VI. Enzymatic conversion of carbon atom 8 of guanosine triphosphate to formic acid. Biochim Biophys Acta. 1966 Mar 28;117(1):275–278. doi: 10.1016/0304-4165(66)90180-2. [DOI] [PubMed] [Google Scholar]
  5. Cha K. W., Jacobson K. B., Yim J. J. Isolation and characterization of GTP cyclohydrolase I from mouse liver. Comparison of normal and the hph-1 mutant. J Biol Chem. 1991 Jul 5;266(19):12294–12300. [PubMed] [Google Scholar]
  6. Cone J. E., Plowman J., Guroff G. Purification and properties of guanosine triphosphate cyclohydrolase and a stimulating protein from Comamonas sp. (ATCC 11299a). J Biol Chem. 1974 Sep 10;249(17):5551–5558. [PubMed] [Google Scholar]
  7. Dalal F. R., Gots J. S. Requirement of GTP for pteridine synthesis in Salmonella typhimurium and its inhibition by AMP. Biochem Biophys Res Commun. 1965 Aug 16;20(4):509–514. doi: 10.1016/0006-291x(65)90609-1. [DOI] [PubMed] [Google Scholar]
  8. Gollnick P., Ishino S., Kuroda M. I., Henner D. J., Yanofsky C. The mtr locus is a two-gene operon required for transcription attenuation in the trp operon of Bacillus subtilis. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8726–8730. doi: 10.1073/pnas.87.22.8726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gray J. V., Golinelli-Pimpaneau B., Knowles J. R. Monofunctional chorismate mutase from Bacillus subtilis: purification of the protein, molecular cloning of the gene, and overexpression of the gene product in Escherichia coli. Biochemistry. 1990 Jan 16;29(2):376–383. doi: 10.1021/bi00454a011. [DOI] [PubMed] [Google Scholar]
  10. Guroff G., Strenkoski C. A. Biosynthesis of pteridines and of phenylalanine hydroxylase cofactor in cell-free extracts of Pseudomonas species (ATCC 11299a). J Biol Chem. 1966 May 25;241(10):2220–2227. [PubMed] [Google Scholar]
  11. Harvey R. J. Growth and initiation of protein synthesis in Escherichia coli in the presence of trimethoprim. J Bacteriol. 1973 Apr;114(1):309–322. doi: 10.1128/jb.114.1.309-322.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hatakeyama K., Harada T., Suzuki S., Watanabe Y., Kagamiyama H. Purification and characterization of rat liver GTP cyclohydrolase I. Cooperative binding of GTP to the enzyme. J Biol Chem. 1989 Dec 25;264(36):21660–21664. [PubMed] [Google Scholar]
  13. Hatakeyama K., Inoue Y., Harada T., Kagamiyama H. Cloning and sequencing of cDNA encoding rat GTP cyclohydrolase I. The first enzyme of the tetrahydrobiopterin biosynthetic pathway. J Biol Chem. 1991 Jan 15;266(2):765–769. [PubMed] [Google Scholar]
  14. Hoch J. A., Nester E. W. Gene-enzyme relationships of aromatic acid biosynthesis in Bacillus subtilis. J Bacteriol. 1973 Oct;116(1):59–66. doi: 10.1128/jb.116.1.59-66.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hoch S. O., Roth C. W., Crawford I. P., Nester E. W. Control of tryptophan biosynthesis by the methyltryptophan resistance gene in Bacillus subtilis. J Bacteriol. 1971 Jan;105(1):38–45. doi: 10.1128/jb.105.1.38-45.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jackson R. J., Shiota T. The nature of the multiple forms of D-erythrodihydroneopterin triphosphate synthetase. Biochim Biophys Acta. 1975 Sep 22;403(1):232–244. doi: 10.1016/0005-2744(75)90025-x. [DOI] [PubMed] [Google Scholar]
  18. Kane J. F. Regulation of a common amidotransferase subunit. J Bacteriol. 1977 Nov;132(2):419–425. doi: 10.1128/jb.132.2.419-425.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Katzenmeier G., Schmid C., Bacher A. Cloning and expression of the putative gene coding for GTP cyclohydrolase I from Escherichia coli. FEMS Microbiol Lett. 1990 Jan 1;54(1-3):231–234. doi: 10.1016/0378-1097(90)90288-2. [DOI] [PubMed] [Google Scholar]
  20. Kuroda M. I., Henner D., Yanofsky C. cis-acting sites in the transcript of the Bacillus subtilis trp operon regulate expression of the operon. J Bacteriol. 1988 Jul;170(7):3080–3088. doi: 10.1128/jb.170.7.3080-3088.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Levenberg B., Kaczmarek D. K. Enzymic release of carbon atom 8 from guanosine triphosphate, an early reaction in the conversion of purines to pteridines. Biochim Biophys Acta. 1966 Mar 28;117(1):272–275. doi: 10.1016/0304-4165(66)90179-6. [DOI] [PubMed] [Google Scholar]
  23. Lovett P. S., Keggins K. M. Bacillus subtilis as a host for molecular cloning. Methods Enzymol. 1979;68:342–357. doi: 10.1016/0076-6879(79)68025-4. [DOI] [PubMed] [Google Scholar]
  24. McLean J. R., Boswell R., O'Donnell J. Cloning and molecular characterization of a metabolic gene with development functions in Drosophila. I. Analysis of the head function of Punch. Genetics. 1990 Dec;126(4):1007–1019. doi: 10.1093/genetics/126.4.1007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schoedon G., Redweik U., Curtius H. C. Purification of GTP cyclohydrolase I from human liver and production of specific monoclonal antibodies. Eur J Biochem. 1989 Jan 2;178(3):627–634. doi: 10.1111/j.1432-1033.1989.tb14491.x. [DOI] [PubMed] [Google Scholar]
  26. Shimotsu H., Henner D. J. Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis. Gene. 1986;43(1-2):85–94. doi: 10.1016/0378-1119(86)90011-9. [DOI] [PubMed] [Google Scholar]
  27. Shimotsu H., Kuroda M. I., Yanofsky C., Henner D. J. Novel form of transcription attenuation regulates expression the Bacillus subtilis tryptophan operon. J Bacteriol. 1986 May;166(2):461–471. doi: 10.1128/jb.166.2.461-471.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Slock J., Stahly D. P., Han C. Y., Six E. W., Crawford I. P. An apparent Bacillus subtilis folic acid biosynthetic operon containing pab, an amphibolic trpG gene, a third gene required for synthesis of para-aminobenzoic acid, and the dihydropteroate synthase gene. J Bacteriol. 1990 Dec;172(12):7211–7226. doi: 10.1128/jb.172.12.7211-7226.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Suzuki Y., Yasui T., Abe S. Occurrence of GTP cyclohydrolase I in Bacillus stearothermophilus. J Biochem. 1979 Dec;86(6):1679–1685. doi: 10.1093/oxfordjournals.jbchem.a132688. [DOI] [PubMed] [Google Scholar]
  30. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Webster K. R., Adari H. Y., Spicer E. K. Bacteriophage T4 regA protein binds to the Shine-Dalgarno region of gene 44 mRNA. Nucleic Acids Res. 1989 Dec 11;17(23):10047–10068. doi: 10.1093/nar/17.23.10047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weisberg E. P., O'Donnell J. M. Purification and characterization of GTP cyclohydrolase I from Drosophila melanogaster. J Biol Chem. 1986 Jan 25;261(3):1453–1458. [PubMed] [Google Scholar]
  33. Yim J. J., Brown G. M. Characteristics of guanosine triphosphate cyclohydrolase I purified from Escherichia coli. J Biol Chem. 1976 Aug 25;251(16):5087–5094. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES