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. 1993 Jun;175(12):3863–3875. doi: 10.1128/jb.175.12.3863-3875.1993

Regulation of the Bacillus subtilis alsS, alsD, and alsR genes involved in post-exponential-phase production of acetoin.

M C Renna 1, N Najimudin 1, L R Winik 1, S A Zahler 1
PMCID: PMC204803  PMID: 7685336

Abstract

Acetoin is a major extracellular product of Bacillus subtilis grown on glucose and other fermentable carbon sources. The enzymes responsible for the formation of acetoin, acetolactate synthase, and acetolactate decarboxylase are synthesized in detectable amounts only in cells that have reached stationary phase. We have cloned and sequenced the genes encoding these enzymes, alsS and alsD, as well as a gene, alsR, that regulates their expression. alsS and alsD appear to compose a single operon, while alsR is transcribed divergently from the alsSD operon. AlsR shows significant homology to the LysR family of bacterial activator proteins, and when alsR is disrupted the alsSD operon is not expressed. Transcriptional fusions to alsS and alsR revealed that AlsR is required for the transcription of the alsSD operon, which increases during stationary phase. Two mutations that cause increased expression of the alsSD operon have been isolated, cloned, and sequenced. They each change an amino acid in the AlsR protein.

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  1. Bohannon D. E., Sonenshein A. L. Positive regulation of glutamate biosynthesis in Bacillus subtilis. J Bacteriol. 1989 Sep;171(9):4718–4727. doi: 10.1128/jb.171.9.4718-4727.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Diderichsen B., Wedsted U., Hedegaard L., Jensen B. R., Sjøholm C. Cloning of aldB, which encodes alpha-acetolactate decarboxylase, an exoenzyme from Bacillus brevis. J Bacteriol. 1990 Aug;172(8):4315–4321. doi: 10.1128/jb.172.8.4315-4321.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dodd I. B., Egan J. B. Systematic method for the detection of potential lambda Cro-like DNA-binding regions in proteins. J Mol Biol. 1987 Apr 5;194(3):557–564. doi: 10.1016/0022-2836(87)90681-4. [DOI] [PubMed] [Google Scholar]
  4. Grandoni J. A., Zahler S. A., Calvo J. M. Transcriptional regulation of the ilv-leu operon of Bacillus subtilis. J Bacteriol. 1992 May;174(10):3212–3219. doi: 10.1128/jb.174.10.3212-3219.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Habeeb L. F., Wang L., Winans S. C. Transcription of the octopine catabolism operon of the Agrobacterium tumor-inducing plasmid pTiA6 is activated by a LysR-type regulatory protein. Mol Plant Microbe Interact. 1991 Jul-Aug;4(4):379–385. doi: 10.1094/mpmi-4-379. [DOI] [PubMed] [Google Scholar]
  6. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  7. Hawkins C. F., Borges A., Perham R. N. A common structural motif in thiamin pyrophosphate-binding enzymes. FEBS Lett. 1989 Sep 11;255(1):77–82. doi: 10.1016/0014-5793(89)81064-6. [DOI] [PubMed] [Google Scholar]
  8. Henikoff S., Haughn G. W., Calvo J. M., Wallace J. C. A large family of bacterial activator proteins. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6602–6606. doi: 10.1073/pnas.85.18.6602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Henkin T. M., Grundy F. J., Nicholson W. L., Chambliss G. H. Catabolite repression of alpha-amylase gene expression in Bacillus subtilis involves a trans-acting gene product homologous to the Escherichia coli lacl and galR repressors. Mol Microbiol. 1991 Mar;5(3):575–584. doi: 10.1111/j.1365-2958.1991.tb00728.x. [DOI] [PubMed] [Google Scholar]
  10. Holtzclaw W. D., Chapman L. F. Degradative acetolactate synthase of Bacillus subtilis: purification and properties. J Bacteriol. 1975 Mar;121(3):917–922. doi: 10.1128/jb.121.3.917-922.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hong G. F., Burn J. E., Johnston A. W. Evidence that DNA involved in the expression of nodulation (nod) genes in Rhizobium binds to the product of the regulatory gene nodD. Nucleic Acids Res. 1987 Dec 10;15(23):9677–9690. doi: 10.1093/nar/15.23.9677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Horvath B., Bachem C. W., Schell J., Kondorosi A. Host-specific regulation of nodulation genes in Rhizobium is mediated by a plant-signal, interacting with the nodD gene product. EMBO J. 1987 Apr;6(4):841–848. doi: 10.1002/j.1460-2075.1987.tb04829.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Huseby N. E., Christensen T. B., Olsen B. R., Stormer F. C. The pH 6 acetolactate-forming enzyme from Aerobacter aerogenes. Subunit structure. Eur J Biochem. 1971 May 28;20(2):209–214. doi: 10.1111/j.1432-1033.1971.tb01381.x. [DOI] [PubMed] [Google Scholar]
  14. Levine M., Epstein S. S., Vaughn R. H. Differential Reactions in the Colon Group of Bacteria. Am J Public Health Nations Health. 1934 May;24(5):505–510. doi: 10.2105/ajph.24.5.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Loken J. P., Stormer F. C. Acetolactate decarboxylase from Aerobacter aerogenes. Purification and properties. Eur J Biochem. 1970 May 1;14(1):133–137. doi: 10.1111/j.1432-1033.1970.tb00270.x. [DOI] [PubMed] [Google Scholar]
  16. Malthe-Sorenssen D., Stormer F. C. The pH 6 acetolactate-forming enzyme from Serratia marcescens. Purification and properties. Eur J Biochem. 1970 May 1;14(1):127–132. doi: 10.1111/j.1432-1033.1970.tb00269.x. [DOI] [PubMed] [Google Scholar]
  17. Moran C. P., Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982;186(3):339–346. doi: 10.1007/BF00729452. [DOI] [PubMed] [Google Scholar]
  18. Nicholson W. L., Park Y. K., Henkin T. M., Won M., Weickert M. J., Gaskell J. A., Chambliss G. H. Catabolite repression-resistant mutations of the Bacillus subtilis alpha-amylase promoter affect transcription levels and are in an operator-like sequence. J Mol Biol. 1987 Dec 20;198(4):609–618. doi: 10.1016/0022-2836(87)90204-x. [DOI] [PubMed] [Google Scholar]
  19. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sone H., Fujii T., Kondo K., Shimizu F., Tanaka J., Inoue T. Nucleotide sequence and expression of the Enterobacter aerogenes alpha-acetolactate decarboxylase gene in brewer's yeast. Appl Environ Microbiol. 1988 Jan;54(1):38–42. doi: 10.1128/aem.54.1.38-42.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stragier P., Patte J. C. Regulation of diaminopimelate decarboxylase synthesis in Escherichia coli. III. Nucleotide sequence and regulation of the lysR gene. J Mol Biol. 1983 Aug 5;168(2):333–350. doi: 10.1016/s0022-2836(83)80022-9. [DOI] [PubMed] [Google Scholar]
  23. Tsau J. L., Guffanti A. A., Montville T. J. Conversion of Pyruvate to Acetoin Helps To Maintain pH Homeostasis in Lactobacillus plantarum. Appl Environ Microbiol. 1992 Mar;58(3):891–894. doi: 10.1128/aem.58.3.891-894.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Vandeyar M. A., Zahler S. A. Chromosomal insertions of Tn917 in Bacillus subtilis. J Bacteriol. 1986 Aug;167(2):530–534. doi: 10.1128/jb.167.2.530-534.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wang L., Helmann J. D., Winans S. C. The A. tumefaciens transcriptional activator OccR causes a bend at a target promoter, which is partially relaxed by a plant tumor metabolite. Cell. 1992 May 15;69(4):659–667. doi: 10.1016/0092-8674(92)90229-6. [DOI] [PubMed] [Google Scholar]
  26. Ward J. B., Jr, Zahler S. A. Genetic studies of leucine biosynthesis in Bacillus subtilis. J Bacteriol. 1973 Nov;116(2):719–726. doi: 10.1128/jb.116.2.719-726.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Weickert M. J., Chambliss G. H. Genetic analysis of the promoter region of the Bacillus subtilis alpha-amylase gene. J Bacteriol. 1989 Jul;171(7):3656–3666. doi: 10.1128/jb.171.7.3656-3666.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wek R. C., Hatfield G. W. Nucleotide sequence and in vivo expression of the ilvY and ilvC genes in Escherichia coli K12. Transcription from divergent overlapping promoters. J Biol Chem. 1986 Feb 15;261(5):2441–2450. [PubMed] [Google Scholar]
  29. Wek R. C., Hatfield G. W. Transcriptional activation at adjacent operators in the divergent-overlapping ilvY and ilvC promoters of Escherichia coli. J Mol Biol. 1988 Oct 5;203(3):643–663. doi: 10.1016/0022-2836(88)90199-4. [DOI] [PubMed] [Google Scholar]
  30. Wek R. C., Hauser C. A., Hatfield G. W. The nucleotide sequence of the ilvBN operon of Escherichia coli: sequence homologies of the acetohydroxy acid synthase isozymes. Nucleic Acids Res. 1985 Jun 11;13(11):3995–4010. doi: 10.1093/nar/13.11.3995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yamazaki H., Ohmura K., Nakayama A., Takeichi Y., Otozai K., Yamasaki M., Tamura G., Yamane K. Alpha-amylase genes (amyR2 and amyE+) from an alpha-amylase-hyperproducing Bacillus subtilis strain: molecular cloning and nucleotide sequences. J Bacteriol. 1983 Oct;156(1):327–337. doi: 10.1128/jb.156.1.327-337.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  33. Yasbin R. E., Wilson G. A., Young F. E. Transformation and transfection in lysogenic strains of Bacillus subtilis: evidence for selective induction of prophage in competent cells. J Bacteriol. 1975 Jan;121(1):296–304. doi: 10.1128/jb.121.1.296-304.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Youngman P., Perkins J. B., Losick R. A novel method for the rapid cloning in Escherichia coli of Bacillus subtilis chromosomal DNA adjacent to Tn917 insertions. Mol Gen Genet. 1984;195(3):424–433. doi: 10.1007/BF00341443. [DOI] [PubMed] [Google Scholar]

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