1887

Abstract

The folate operon contains nine genes. The first six genes are involved in the biosynthesis of folic acid and tryptophan and have been characterized previously. The 3-region of the folate operon contains three additional ORFs: , potentially encoding a DNA-binding protein of 68 amino acids, , encoding a protein of 338 amino acids with homology to the Orf1 of the operon, and a putative lysyl-tRNA synthetase gene (). Four transcripts were identified which encode the first two, eight or all nine proteins or only the last protein LysS. The folate operon contains two promoters, one upstream of the first gene and the second preceding . Transcription of the entire folate operon starts 33 bp upstream of the ATG codon of , the first gene of the operon. The -encoded RNA-binding attenuation protein (TRAP) dramatically reduces the steady-state levels of the folate operon transcripts encoding the first eight and all nine proteins, but only has a relatively small effect on the steady-state level of the 2.1 kb transcript encoding the first two genes of the operon, and . In addition, transcription of the folate operon is regulated in a growth-phase-dependent manner. Transcripts were present in very low levels after mid-exponential phase, but were dramatically increased directly after transfer of the cells to fresh medium. These results indicate that transcription of the folate operon is regulated by TRAP and also depends on the growth phase of the culture.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-3-979
1997-03-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/3/mic-143-3-979.html?itemId=/content/journal/micro/10.1099/00221287-143-3-979&mimeType=html&fmt=ahah

References

  1. Babitzke P., Yanofsky C. 1993; Reconstitution of Bacillus subtilis trp attenuation in vitro with TRAP, the trp RNA-binding attenuation protein.. Proc Natl Acad Sci USA 90133–137
    [Google Scholar]
  2. Babitzke P., Gollnick P., Yanofsky C. 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.. J Bacteriol 174:2059–2064
    [Google Scholar]
  3. Babitzke P., Stults J.T., Shire S.J., Yanofsky C. 1994; TRAP, the trp RNA-binding attenuation protein of Bacillus subtilis, is a multisubunit complex that appears to recognize G/UAG repeats in the trpEDCFBA and trpG transcripts.. J Biol Chem 269:16597–16604
    [Google Scholar]
  4. Babitzke P., Yealy J., Campanelli D. 1996; Interaction of the trp RNA-binding attenuation protein (TRAP) of Bacillus subtilis with RNA: effects of the number of GAG repeats, the nucleotides separating adjacent repeats, and secondary structures.. J Bacteriol 178:5159–5163
    [Google Scholar]
  5. Béjar S., Cam K., Bouché J.-P. 1986; Control of cell division in Escherichia coli. DNA sequence of dicA and of a second gene complementing mutation dicAl, dicC.. Nucleic Acids Res 14:6821–6833
    [Google Scholar]
  6. Belyaeva T., Griffiths L., Minchin S., Cole J., Busby S. 1993; The Escherichia coli cysG promoter belongs to the ‘extended’ 10’ class of bacterial promoters.. Biochem J 296:851–857
    [Google Scholar]
  7. Brown G.M., Williamson M. 1982; Biosynthesis of riboflavin, folic acid, thiamine, and pantothenic acid.. Adv Enzymol Relat Areas Mol Biol 53:345–381
    [Google Scholar]
  8. Foster-Hartnett D., Cullen P.J., Gabbert K.K., Kranz R.G. 1993; Sequence, genetic, and lacZ fusion analyses of a nifR3-ntrB-ntrC operon in Rhodobacter capsulatus.. Mol Microbiol 8:903–914
    [Google Scholar]
  9. Gallant J.A. 1979; Stringent control in E. coli.. Annu Rev Genet 13:393–415
    [Google Scholar]
  10. Gaur N.K., Oppenheim J., Smith I. 1991; The Bacillus subtilis sin gene, a regulator of alternate developmental processes, codes for a DNA-binding protein.. J Bacteriol 173:678–686
    [Google Scholar]
  11. Gausing K. 1977; Regulation of ribosome production in Escherichia coli: synthesis and stability of ribosomal RNA and of ribosomal protein messenger RNA at different growth rates.. J Mol Biol 115:335–354
    [Google Scholar]
  12. Goncharoff P., Nichols B.P. 1984; Nucleotide sequence of E. coli pabB indicates a common evolutionary origin of p-aminobenzoate synthase and anthranilate synthase.. J Bacteriol 159:57–62
    [Google Scholar]
  13. Grundy F.J., Henkin T.M. 1993; tRNA as a positive regulator of transcription antitermination in B. subtilis.. Cell 74:475–482
    [Google Scholar]
  14. Helmann J.D. 1995; Compilation and analysis of Bacillus subtilis σA-dependent promoter sequences: evidence for extended contact between RNA polymerase and upstream promoter DNA.. Nucleic Acids Res 23:2351–2360
    [Google Scholar]
  15. van Kaer L., van Montagu M., Dhaese P. 1987; Transcriptional control in the EcoRI-F immunity region of Bacillus subtilis phage ɸ105.. J Mol Biol 197:55–67
    [Google Scholar]
  16. Kane J.F., Holmes W.M., Jensen R.A. 1972; Metabolic interlock: the dual function of folate pathway gene as an extra-operonic gene of tryptophan biosynthesis.. J Biol Chem 247:1587–1596
    [Google Scholar]
  17. Kuroda M.I., Henner D., Yanofsky C. 1988; cis-Acting sites in the transcript of the Bacillus subtilis trp operon regulate the expression of the operon.. J Bacteriol 170:3080–3088
    [Google Scholar]
  18. Lacks S.A., Greenberg B., Lopez P. 1995; A cluster of four genes encoding enzymes for five steps in the folate biosynthetic pathway of Streptococcus pneumoniae.. J Bacteriol 177:66–74
    [Google Scholar]
  19. Lopez P., Lacks S.A. 1993; A bifunctional protein in the folate biosynthetic pathway of Streptococcus pneumoniae with dihydroneopterin aldolase and hydroxymethyldihydropterin pyro-phosphokinase activities.. J Bacteriol 175:2214–2220
    [Google Scholar]
  20. McDonald K.O., Burke W.F. 1982; Cloning of the Bacillus subtilis sulfanilamide resistance gene in Bacillus subtilis.. J Bacteriol 149:391–394
    [Google Scholar]
  21. Maes M., Messens E. 1992; Phenol as grinding material in RNA preparations.. Nucleic Acids Res 20:4374
    [Google Scholar]
  22. Margolis P.S., Driks A., Losick R. 1993; Sporulation gene spoIlB from Bacillus subtilis.. J Bacteriol 175:528–540
    [Google Scholar]
  23. Melton D.A., Krieg P.A., Rebagliati M.R., Maniatis T., Zinn K., Green M.R. 1984; Efficient in vitro synthesis of biologically active RNA and RNA hybridisation probes from plasmids containing a bacteriophage SP6 promoter.. Nucleic Acids Res 12:7035–7056
    [Google Scholar]
  24. Miller J.H. 1972 In Experiments in Molecular Genetics pp. 353–355 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  25. Nakamura Y., loto K. 1993; Control and function of lysyl-tRNA synthetases: diversity and co-ordination.. Mol Microbiol 10:225–231
    [Google Scholar]
  26. Nakano M.M., Xia L., Zuber P. 1991; Transcription initiation region of the srfA operon, which is controlled by the comP-comA signal transduction system in Bacillus subtilis.. J Bacteriol 173:5487–5493
    [Google Scholar]
  27. Ninnemann O., Koch C., Kahmann R. 1992; The Escherichia coli fis promoter is subject to stringent control and autoregulation.. EMBO J 11:1075–1083
    [Google Scholar]
  28. Ogasawara N., Nakai S., Yoshikawa H. 1994; Systematic sequencing of the 180 kilobase region of the Bacillus subtilis chromosome containing the replication origin.. DNA Res 1:1–14
    [Google Scholar]
  29. Otridge G., Gollnick P. 1993; MtrB from Bacillus subtilis binds specifically to trp leader RNA in a tryptophan-dependent manner. Proc Natl Acad Sci USA 90:128–132
    [Google Scholar]
  30. Patriarca E.J., Riccio A., Tate R., Colonna-Romaro S., Laccarino M., Defez R. 1993; The ntrBC genes of Rhizobium leguminosarum are part of a complex operon subject to negative regulation.. Mol Microbiol 9:569–577
    [Google Scholar]
  31. Piggot P.J., Curtis C.A.M., Hoch J.A. 1984; Use of integrational plasmid vectors to demonstrate the polycistronic nature of a polycistronic unit (spoIIA) required for sporulation of Bacillus subtilis.. J Gen Microbiol 130:2123–2136
    [Google Scholar]
  32. Racine F.M., Steinberg W. 1974; Genetic location of two mutations affecting the lysyl-transfer ribonucleic acid synthetase of Bacillus subtilis.. J Bacteriol 120:384–389
    [Google Scholar]
  33. Saito H., Shibata T., Ando T. 1979; Mapping of genes determining nonpermissiveness and host-specific restriction to bacteriophages in Bacillus subtilis Marburg.. Mol Gen Genet 170:117–122
    [Google Scholar]
  34. de Saizieu A., Vankan P., van Loon A.P.G.M. 1995; Enzymatic characterisation of Bacillus subtilis GTP cyclohydrolase I. Evidence for a chemical dephosphorylation of dihydro-neopterin triphosphate.. Biocbem J 306:371–377
    [Google Scholar]
  35. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Elarbor, NY: Cold Spring Elarbor Laboratory;
    [Google Scholar]
  36. Simpson W.J., LaPenta D., Chen C., Cleary P.P. 1990; Coregulation of type 12 M protein and streptococcal C5a peptidase genes in group A streptococci: evidence for a virulence regulon controlled by the virR locus.. J Bacteriol 172:696–700
    [Google Scholar]
  37. Slock J., Stahly D.P., Han C.Y., Six E.W., Crawford I.P. 1990; An apparent Bacillus subtilis folic acid biosynthetic operon containing pab, an amphibolic trpG gene, a third gene required for synthesis of ptfra'-aminobenzoic acid and the dihydropteroate synthase gene. . J Bacteriol 172:7211–7226
    [Google Scholar]
  38. Sokawa J., Sokawa Y. 1978; Relaxation effect of chloramphenicol on the stringent control in Escherichia coli.. J Biocbem 83:1699–1705
    [Google Scholar]
  39. Strauch M.A., Spiegelman G.P., Perego M., Johnson W.C., Burbulys D., Hoch J.A. 1989; The transition state transcription regulator abrB of Bacillus subtilis is a DNA-binding protein.. EMBO J 8:1615–1621
    [Google Scholar]
  40. Stuber D., Matile H., Garotta G. 1990; System for high-level production in Escherichia coli and rapid purification of recombinant proteins: application to epitope mapping, preparation of antibodies, and structure-function analysis.. In Immunological Methods 4 pp. 121–152 Edited by Lefkovits I., Pernis B. Orlando, FL: Academic Press;
    [Google Scholar]
  41. Turner R.J., Lu Y., Switzer R.L. 1994; Regulation of the Bacillus subtilis pyrimidine biosynthetic (pyr) gene cluster by an autogenous transcriptional attenuation mechanism.. J Bacteriol 176:3708–3722
    [Google Scholar]
  42. Yang M., de Saizieu A., van Loon A.P.G.M., Gollnick P. 1995; Translation of trpG in Bacillus subtilis is regulated by the trp RNA-binding attenuation protein (TRAP).. J Bacteriol 177:4272–4278
    [Google Scholar]
  43. Zuber P., Losick R. 1983; Use of a lacZ fusion to study the role of the spoO genes of Bacillus subtilis in developmental regulation.. Cell 35:275–283
    [Google Scholar]
  44. Zuber P., Losick R. 1987; Role of AbrB in SpoOA- and SpoOB- dependent utilization of a sporulation promoter in Bacillus subtilis.. J Bacteriol 169:2223–2230
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-3-979
Loading
/content/journal/micro/10.1099/00221287-143-3-979
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error