Abstract
We report the cloning and characterization of a cell division gene, herein designated divIC, from the gram-positive, spore-forming bacterium Bacillus subtilis. This gene was previously identified on the basis of a temperature-sensitive mutation, div-355, that blocks septum formation at restrictive temperatures. We show that the divIC gene is a 125-codon open reading frame that is capable of encoding a protein of 14.7 kDa and that div-355 is a 5-bp duplication near the 3' end of the open reading frame. We also show that divIC is an essential gene by use of an in vitro-constructed null mutation. In confirmation and extension of earlier results, we show that divIC is necessary for both vegetative and sporulation septum formation, and we demonstrate that it is required for the activation of genes expressed under the control of the sporulation transcription factors sigma F and sigma E. The divIC gene is located 1.3 kb upstream of the coding sequence for the sporulation gene spoIIE. Between divIC and spoIIE is a 128-codon open reading frame whose predicted product contains a region of similarity to the RNA-binding domains of polynucleotide phosphorylase and ribosomal protein S1 from Escherichia coli and two putative tRNA genes for methionyl-tRNA and glutamyl-tRNA, the gene order being divIC orf128 tRNA(Met) tRNA(Glu) spoIIE.
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- Alber T. Structure of the leucine zipper. Curr Opin Genet Dev. 1992 Apr;2(2):205–210. doi: 10.1016/s0959-437x(05)80275-8. [DOI] [PubMed] [Google Scholar]
- Beall B., Lowe M., Lutkenhaus J. Cloning and characterization of Bacillus subtilis homologs of Escherichia coli cell division genes ftsZ and ftsA. J Bacteriol. 1988 Oct;170(10):4855–4864. doi: 10.1128/jb.170.10.4855-4864.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Beall B., Lutkenhaus J. Nucleotide sequence and insertional inactivation of a Bacillus subtilis gene that affects cell division, sporulation, and temperature sensitivity. J Bacteriol. 1989 Dec;171(12):6821–6834. doi: 10.1128/jb.171.12.6821-6834.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bi E. F., Lutkenhaus J. FtsZ ring structure associated with division in Escherichia coli. Nature. 1991 Nov 14;354(6349):161–164. doi: 10.1038/354161a0. [DOI] [PubMed] [Google Scholar]
- Bilofsky H. S., Burks C. The GenBank genetic sequence data bank. Nucleic Acids Res. 1988 Mar 11;16(5):1861–1863. doi: 10.1093/nar/16.5.1861. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Callister H., Wake R. G. Characterization and mapping of temperature-sensitive division initiation mutations of Bacillus subtilis. J Bacteriol. 1981 Feb;145(2):1042–1051. doi: 10.1128/jb.145.2.1042-1051.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clary D. O., Griff I. C., Rothman J. E. SNAPs, a family of NSF attachment proteins involved in intracellular membrane fusion in animals and yeast. Cell. 1990 May 18;61(4):709–721. doi: 10.1016/0092-8674(90)90482-t. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- Guzman L. M., Barondess J. J., Beckwith J. FtsL, an essential cytoplasmic membrane protein involved in cell division in Escherichia coli. J Bacteriol. 1992 Dec;174(23):7716–7728. [PMC free article] [PubMed] [Google Scholar]
- Guzmán P., Westpheling J., Youngman P. Characterization of the promoter region of the Bacillus subtilis spoIIE operon. J Bacteriol. 1988 Apr;170(4):1598–1609. doi: 10.1128/jb.170.4.1598-1609.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harry E. J., Stewart B. J., Wake R. G. Characterization of mutations in divIB of Bacillus subtilis and cellular localization of the DivIB protein. Mol Microbiol. 1993 Feb;7(4):611–621. doi: 10.1111/j.1365-2958.1993.tb01152.x. [DOI] [PubMed] [Google Scholar]
- Harry E. J., Wake R. G. Cloning and expression of a Bacillus subtilis division initiation gene for which a homolog has not been identified in another organism. J Bacteriol. 1989 Dec;171(12):6835–6839. doi: 10.1128/jb.171.12.6835-6839.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Karmazyn-Campelli C., Fluss L., Leighton T., Stragier P. The spoIIN279(ts) mutation affects the FtsA protein of Bacillus subtilis. Biochimie. 1992 Jul-Aug;74(7-8):689–694. doi: 10.1016/0300-9084(92)90141-z. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Lutkenhaus J. FtsZ ring in bacterial cytokinesis. Mol Microbiol. 1993 Aug;9(3):403–409. doi: 10.1111/j.1365-2958.1993.tb01701.x. [DOI] [PubMed] [Google Scholar]
- Margolis P. S., Driks A., Losick R. Sporulation gene spoIIB from Bacillus subtilis. J Bacteriol. 1993 Jan;175(2):528–540. doi: 10.1128/jb.175.2.528-540.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McKnight S. L. Molecular zippers in gene regulation. Sci Am. 1991 Apr;264(4):54–64. doi: 10.1038/scientificamerican0491-54. [DOI] [PubMed] [Google Scholar]
- 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]
- Mukherjee A., Dai K., Lutkenhaus J. Escherichia coli cell division protein FtsZ is a guanine nucleotide binding protein. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):1053–1057. doi: 10.1073/pnas.90.3.1053. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nelson K. K., Lemmon S. K. Suppressors of clathrin deficiency: overexpression of ubiquitin rescues lethal strains of clathrin-deficient Saccharomyces cerevisiae. Mol Cell Biol. 1993 Jan;13(1):521–532. doi: 10.1128/mcb.13.1.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Okubo S., Yanagida T. Isolation of a suppressor mutant in Bacillus subtilis. J Bacteriol. 1968 Mar;95(3):1187–1188. doi: 10.1128/jb.95.3.1187-1188.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Régnier P., Grunberg-Manago M., Portier C. Nucleotide sequence of the pnp gene of Escherichia coli encoding polynucleotide phosphorylase. Homology of the primary structure of the protein with the RNA-binding domain of ribosomal protein S1. J Biol Chem. 1987 Jan 5;262(1):63–68. [PubMed] [Google Scholar]
- 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]
- 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]
- Schnier J., Kimura M., Foulaki K., Subramanian A. R., Isono K., Wittmann-Liebold B. Primary structure of Escherichia coli ribosomal protein S1 and of its gene rpsA. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1008–1011. doi: 10.1073/pnas.79.4.1008. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Wang X., Lutkenhaus J. The FtsZ protein of Bacillus subtilis is localized at the division site and has GTPase activity that is dependent upon FtsZ concentration. Mol Microbiol. 1993 Aug;9(3):435–442. doi: 10.1111/j.1365-2958.1993.tb01705.x. [DOI] [PubMed] [Google Scholar]
- Yanouri A., Daniel R. A., Errington J., Buchanan C. E. Cloning and sequencing of the cell division gene pbpB, which encodes penicillin-binding protein 2B in Bacillus subtilis. J Bacteriol. 1993 Dec;175(23):7604–7616. doi: 10.1128/jb.175.23.7604-7616.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Young M. Use of temperature-sensitive mutants to study gene expression during sporulation in Bacillus subtilis. J Bacteriol. 1976 May;126(2):928–936. doi: 10.1128/jb.126.2.928-936.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- de Boer P., Crossley R., Rothfield L. The essential bacterial cell-division protein FtsZ is a GTPase. Nature. 1992 Sep 17;359(6392):254–256. doi: 10.1038/359254a0. [DOI] [PubMed] [Google Scholar]

