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. 1997 Mar;179(5):1671–1683. doi: 10.1128/jb.179.5.1671-1683.1997

The divIVA minicell locus of Bacillus subtilis.

J H Cha 1, G C Stewart 1
PMCID: PMC178881  PMID: 9045828

Abstract

The Bacillus subtilis divIVA1 mutation causes misplacement of the septum during cell division, resulting in the formation of small, circular, anucleate minicells. This study reports the cloning and sequence analysis of 2.4 kb of the B. subtilis chromosome including the divIVA locus. Three open reading frames were identified: orf, whose function is unknown; divIVA; and isoleucyl tRNA synthetase (ileS). We identified the point mutation in the divIVA1 mutant allele. Inactivation of divIVA produces a minicell phenotype, whereas overproduction of DivIVA results in a filamentation phenotype. Mutants with mutations at both of the minicell loci of B. subtilis, divIVA and divIVB, possess a minicell phenotype identical to that of the DivIVB- mutant. The DivIVA-mutants, but not the DivIVB- mutants, show a decrease in sporulation efficiency and a delay in the kinetics of endospore formation. The data support a model in which divIVA encodes the topological specificity subunit of the minCD system. The model suggests that DivIVA acts as a pilot protein, directing minCD to the polar septation sites. DivIVA also appears to be the interface between a sporulation component and MinCD, freeing up the polar septation sites for use during the asymmetric septation event of the sporulation process.

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Selected References

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  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. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. 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]
  6. 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]
  7. Butler Y. X., Abhayawardhane Y., Stewart G. C. Amplification of the Bacillus subtilis maf gene results in arrested septum formation. J Bacteriol. 1993 May;175(10):3139–3145. doi: 10.1128/jb.175.10.3139-3145.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Coyne S. I., Mendelson N. H. Clonal analysis of cell division in the Bacillus subtilis div IV-B1 minicell-producing mutant. J Bacteriol. 1974 Apr;118(1):15–20. doi: 10.1128/jb.118.1.15-20.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Erickson R. J., Copeland J. C. Structure and replication of chromosomes in competent cells of Bacillus subtilis. J Bacteriol. 1972 Mar;109(3):1075–1084. doi: 10.1128/jb.109.3.1075-1084.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  13. Grundy F. J., Henkin T. M. Conservation of a transcription antitermination mechanism in aminoacyl-tRNA synthetase and amino acid biosynthesis genes in gram-positive bacteria. J Mol Biol. 1994 Jan 14;235(2):798–804. doi: 10.1006/jmbi.1994.1038. [DOI] [PubMed] [Google Scholar]
  14. Hermans P. W., Abebe F., Kuteyi V. I., Kolk A. H., Thole J. E., Harboe M. Molecular and immunological characterization of the highly conserved antigen 84 from Mycobacterium tuberculosis and Mycobacterium leprae. Infect Immun. 1995 Mar;63(3):954–960. doi: 10.1128/iai.63.3.954-960.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hirota Y., Jacob F., Ryter A., Buttin G., Nakai T. On the process of cellular division in Escherichia coli. I. Asymmetrical cell division and production of deoxyribonucleic acid-less bacteria. J Mol Biol. 1968 Jul 14;35(1):175–192. doi: 10.1016/s0022-2836(68)80046-4. [DOI] [PubMed] [Google Scholar]
  16. Honeyman A. L., Stewart G. C. The nucleotide sequence of the rodC operon of Bacillus subtilis. Mol Microbiol. 1989 Sep;3(9):1257–1268. doi: 10.1111/j.1365-2958.1989.tb00276.x. [DOI] [PubMed] [Google Scholar]
  17. Lee S., Price C. W. The minCD locus of Bacillus subtilis lacks the minE determinant that provides topological specificity to cell division. Mol Microbiol. 1993 Feb;7(4):601–610. doi: 10.1111/j.1365-2958.1993.tb01151.x. [DOI] [PubMed] [Google Scholar]
  18. Levin P. A., Losick R. Characterization of a cell division gene from Bacillus subtilis that is required for vegetative and sporulation septum formation. J Bacteriol. 1994 Mar;176(5):1451–1459. doi: 10.1128/jb.176.5.1451-1459.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Levin P. A., Losick R. Transcription factor Spo0A switches the localization of the cell division protein FtsZ from a medial to a bipolar pattern in Bacillus subtilis. Genes Dev. 1996 Feb 15;10(4):478–488. doi: 10.1101/gad.10.4.478. [DOI] [PubMed] [Google Scholar]
  20. Levin P. A., Margolis P. S., Setlow P., Losick R., Sun D. Identification of Bacillus subtilis genes for septum placement and shape determination. J Bacteriol. 1992 Nov;174(21):6717–6728. doi: 10.1128/jb.174.21.6717-6728.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mendelson N. H., Cole R. M. Genetic regulation of cell division initiation in Bacillus subtilis. J Bacteriol. 1972 Nov;112(2):994–1003. doi: 10.1128/jb.112.2.994-1003.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pichoff S., Vollrath B., Touriol C., Bouché J. P. Deletion analysis of gene minE which encodes the topological specificity factor of cell division in Escherichia coli. Mol Microbiol. 1995 Oct;18(2):321–329. doi: 10.1111/j.1365-2958.1995.mmi_18020321.x. [DOI] [PubMed] [Google Scholar]
  23. Quinn C. L., Stephenson B. T., Switzer R. L. Functional organization and nucleotide sequence of the Bacillus subtilis pyrimidine biosynthetic operon. J Biol Chem. 1991 May 15;266(14):9113–9127. [PubMed] [Google Scholar]
  24. 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]
  25. Rothfield L. I., Zhao C. R. How do bacteria decide where to divide? Cell. 1996 Jan 26;84(2):183–186. doi: 10.1016/s0092-8674(00)80971-x. [DOI] [PubMed] [Google Scholar]
  26. SAITO H., MIURA K. I. PREPARATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID BY PHENOL TREATMENT. Biochim Biophys Acta. 1963 Aug 20;72:619–629. [PubMed] [Google Scholar]
  27. Schaeffer P., Millet J., Aubert J. P. Catabolic repression of bacterial sporulation. Proc Natl Acad Sci U S A. 1965 Sep;54(3):704–711. doi: 10.1073/pnas.54.3.704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Steinmetz M., Richter R. Plasmids designed to alter the antibiotic resistance expressed by insertion mutations in Bacillus subtilis, through in vivo recombination. Gene. 1994 May 3;142(1):79–83. doi: 10.1016/0378-1119(94)90358-1. [DOI] [PubMed] [Google Scholar]
  29. Van Alstyne D., Simon M. I. Division mutants of Bacillus subtilis: isolation and PBS1 transduction of division-specific markers. J Bacteriol. 1971 Dec;108(3):1366–1379. doi: 10.1128/jb.108.3.1366-1379.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. 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]
  33. 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]
  34. Wagner P. M., Stewart G. C. Role and expression of the Bacillus subtilis rodC operon. J Bacteriol. 1991 Jul;173(14):4341–4346. doi: 10.1128/jb.173.14.4341-4346.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ward J. E., Jr, Lutkenhaus J. Overproduction of FtsZ induces minicell formation in E. coli. Cell. 1985 Oct;42(3):941–949. doi: 10.1016/0092-8674(85)90290-9. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Zhao C. R., de Boer P. A., Rothfield L. I. Proper placement of the Escherichia coli division site requires two functions that are associated with different domains of the MinE protein. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4313–4317. doi: 10.1073/pnas.92.10.4313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. de Boer P. A., Crossley R. E., Hand A. R., Rothfield L. I. The MinD protein is a membrane ATPase required for the correct placement of the Escherichia coli division site. EMBO J. 1991 Dec;10(13):4371–4380. doi: 10.1002/j.1460-2075.1991.tb05015.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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]
  40. 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]
  41. de Boer P. A., Crossley R. E., Rothfield L. I. Roles of MinC and MinD in the site-specific septation block mediated by the MinCDE system of Escherichia coli. J Bacteriol. 1992 Jan;174(1):63–70. doi: 10.1128/jb.174.1.63-70.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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