Identification of Bacillus subtilis genes for septum placement and shape determination (original) (raw)

Abstract

The Bacillus subtilis divIVB1 mutation causes aberrant positioning of the septum during cell division, resulting in the formation of small, anucleate cells known as minicells. We report the cloning of the wild-type allele of divIVB1 and show that the mutation lies within a stretch of DNA containing two open reading frames whose predicted products are in part homologous to the products of the Escherichia coli minicell genes minC and minD. Just upstream of minC and minD, and in the same orientation, are three genes whose products are homologous to the products of the E. coli shape-determining genes mreB, mreC, and mreD. The B. subtilis mreB, mreC, and mreD genes are the site of a conditional mutation (rodB1) that causes the production of aberrantly shaped cells under restrictive conditions. Northern (RNA) hybridization experiments and disruption experiments based on the use of integrational plasmids indicate that the mre and min genes constitute a five-cistron operon. The possible involvement of min gene products in the switch from medial to polar placement of the septum during sporulation is discussed.

6717

Images in this article

Selected References

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

  1. Adler H. I., Fisher W. D., Cohen A., Hardigree A. A. MINIATURE escherichia coli CELLS DEFICIENT IN DNA. Proc Natl Acad Sci U S A. 1967 Feb;57(2):321–326. doi: 10.1073/pnas.57.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beall B., Lutkenhaus J. FtsZ in Bacillus subtilis is required for vegetative septation and for asymmetric septation during sporulation. Genes Dev. 1991 Mar;5(3):447–455. doi: 10.1101/gad.5.3.447. [DOI] [PubMed] [Google Scholar]
  3. Beall B., Lutkenhaus J. Impaired cell division and sporulation of a Bacillus subtilis strain with the ftsA gene deleted. J Bacteriol. 1992 Apr;174(7):2398–2403. doi: 10.1128/jb.174.7.2398-2403.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bi E., Lutkenhaus J. Interaction between the min locus and ftsZ. J Bacteriol. 1990 Oct;172(10):5610–5616. doi: 10.1128/jb.172.10.5610-5616.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cutting S., Roels S., Losick R. Sporulation operon spoIVF and the characterization of mutations that uncouple mother-cell from forespore gene expression in Bacillus subtilis. J Mol Biol. 1991 Oct 20;221(4):1237–1256. doi: 10.1016/0022-2836(91)90931-u. [DOI] [PubMed] [Google Scholar]
  6. Davie E., Sydnor K., Rothfield L. I. Genetic basis of minicell formation in Escherichia coli K-12. J Bacteriol. 1984 Jun;158(3):1202–1203. doi: 10.1128/jb.158.3.1202-1203.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Doi M., Wachi M., Ishino F., Tomioka S., Ito M., Sakagami Y., Suzuki A., Matsuhashi M. Determinations of the DNA sequence of the mreB gene and of the gene products of the mre region that function in formation of the rod shape of Escherichia coli cells. J Bacteriol. 1988 Oct;170(10):4619–4624. doi: 10.1128/jb.170.10.4619-4624.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dubnau D., Davidoff-Abelson R. Fate of transforming DNA following uptake by competent Bacillus subtilis. I. Formation and properties of the donor-recipient complex. J Mol Biol. 1971 Mar 14;56(2):209–221. doi: 10.1016/0022-2836(71)90460-8. [DOI] [PubMed] [Google Scholar]
  10. Ellis D. M., Dean D. H. Location of the Bacillus subtilis temperate bacteriophage phi 105 attP attachment site. J Virol. 1986 Apr;58(1):223–224. doi: 10.1128/jvi.58.1.223-224.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fort P., Errington J. Nucleotide sequence and complementation analysis of a polycistronic sporulation operon, spoVA, in Bacillus subtilis. J Gen Microbiol. 1985 May;131(5):1091–1105. doi: 10.1099/00221287-131-5-1091. [DOI] [PubMed] [Google Scholar]
  12. Henner D. J., Hoch J. A. The Bacillus subtilis chromosome. Microbiol Rev. 1980 Mar;44(1):57–82. doi: 10.1128/mr.44.1.57-82.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hitchins A. D., Slepecky R. A. Bacterial sporulation as a modified procaryotic cell division. Nature. 1969 Aug 23;223(5208):804–807. doi: 10.1038/223804a0. [DOI] [PubMed] [Google Scholar]
  14. Hoch J. A. Genetic analysis of pleiotropic negative sporulation mutants in Bacillus subtilis. J Bacteriol. 1971 Mar;105(3):896–901. doi: 10.1128/jb.105.3.896-901.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jaffé A., Boye E., D'Ari R. Rule governing the division pattern in Escherichia coli minB and wild-type filaments. J Bacteriol. 1990 Jun;172(6):3500–3502. doi: 10.1128/jb.172.6.3500-3502.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Karamata D., McConnell M., Rogers H. J. Mapping of rod mutants of Bacillus subtilis. J Bacteriol. 1972 Jul;111(1):73–79. doi: 10.1128/jb.111.1.73-79.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Labie C., Bouché F., Bouché J. P. Minicell-forming mutants of Escherichia coli: suppression of both DicB- and MinD-dependent division inhibition by inactivation of the minC gene product. J Bacteriol. 1990 Oct;172(10):5852–5855. doi: 10.1128/jb.172.10.5852-5855.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lamont I. L., Mandelstam J. Identification of a new sporulation locus, spoIIIF, in Bacillus subtilis. J Gen Microbiol. 1984 May;130(5):1253–1261. doi: 10.1099/00221287-130-5-1253. [DOI] [PubMed] [Google Scholar]
  19. Marrero R., Yasbin R. E. Evidence for circular permutation of the prophage genome of Bacillus subtilis bacteriophage phi 105. J Virol. 1986 Mar;57(3):1145–1148. doi: 10.1128/jvi.57.3.1145-1148.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Mulder E., Woldringh C. L., Tétart F., Bouché J. P. New minC mutations suggest different interactions of the same region of division inhibitor MinC with proteins specific for minD and dicB coinhibition pathways. J Bacteriol. 1992 Jan;174(1):35–39. doi: 10.1128/jb.174.1.35-39.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Reeve J. N., Mendelson N. H., Coyne S. I., Hallock L. L., Cole R. M. Minicells of Bacillus subtilis. J Bacteriol. 1973 May;114(2):860–873. doi: 10.1128/jb.114.2.860-873.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Roels S., Driks A., Losick R. Characterization of spoIVA, a sporulation gene involved in coat morphogenesis in Bacillus subtilis. J Bacteriol. 1992 Jan;174(2):575–585. doi: 10.1128/jb.174.2.575-585.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rogers H. J., McConnell M., Burdett I. D. The isolation and characterization of mutants of Bacillus subtilis and Bacillus licheniformis with disturbed morphology and cell division. J Gen Microbiol. 1970 May;61(2):155–171. doi: 10.1099/00221287-61-2-155. [DOI] [PubMed] [Google Scholar]
  25. Sandman K., Losick R., Youngman P. Genetic analysis of Bacillus subtilis spo mutations generated by Tn917-mediated insertional mutagenesis. Genetics. 1987 Dec;117(4):603–617. doi: 10.1093/genetics/117.4.603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Varley A. W., Stewart G. C. The divIVB region of the Bacillus subtilis chromosome encodes homologs of Escherichia coli septum placement (minCD) and cell shape (mreBCD) determinants. J Bacteriol. 1992 Nov;174(21):6729–6742. doi: 10.1128/jb.174.21.6729-6742.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wachi M., Doi M., Okada Y., Matsuhashi M. New mre genes mreC and mreD, responsible for formation of the rod shape of Escherichia coli cells. J Bacteriol. 1989 Dec;171(12):6511–6516. doi: 10.1128/jb.171.12.6511-6516.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wachi M., Doi M., Tamaki S., Park W., Nakajima-Iijima S., Matsuhashi M. Mutant isolation and molecular cloning of mre genes, which determine cell shape, sensitivity to mecillinam, and amount of penicillin-binding proteins in Escherichia coli. J Bacteriol. 1987 Nov;169(11):4935–4940. doi: 10.1128/jb.169.11.4935-4940.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wachi M., Doi M., Ueda T., Ueki M., Tsuritani K., Nagai K., Matsuhashi M. Sequence of the downstream flanking region of the shape-determining genes mreBCD of Escherichia coli. Gene. 1991 Sep 30;106(1):135–136. doi: 10.1016/0378-1119(91)90578-y. [DOI] [PubMed] [Google Scholar]
  31. Wachi M., Matsuhashi M. Negative control of cell division by mreB, a gene that functions in determining the rod shape of Escherichia coli cells. J Bacteriol. 1989 Jun;171(6):3123–3127. doi: 10.1128/jb.171.6.3123-3127.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. de Boer P. A., Cook W. R., Rothfield L. I. Bacterial cell division. Annu Rev Genet. 1990;24:249–274. doi: 10.1146/annurev.ge.24.120190.001341. [DOI] [PubMed] [Google Scholar]
  34. de Boer P. A., Crossley R. E., Rothfield L. I. A division inhibitor and a topological specificity factor coded for by the minicell locus determine proper placement of the division septum in E. coli. Cell. 1989 Feb 24;56(4):641–649. doi: 10.1016/0092-8674(89)90586-2. [DOI] [PubMed] [Google Scholar]
  35. de Boer P. A., Crossley R. E., Rothfield L. I. Central role for the Escherichia coli minC gene product in two different cell division-inhibition systems. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1129–1133. doi: 10.1073/pnas.87.3.1129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. de Boer P. A., Crossley R. E., Rothfield L. I. Isolation and properties of minB, a complex genetic locus involved in correct placement of the division site in Escherichia coli. J Bacteriol. 1988 May;170(5):2106–2112. doi: 10.1128/jb.170.5.2106-2112.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]