Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Jan;174(2):477–485. doi: 10.1128/jb.174.2.477-485.1992

Temporal expression of a membrane-associated protein putatively involved in repression of initiation of DNA replication in Bacillus subtilis.

B Eident-Wilkinson 1, L Mele 1, J Laffan 1, W Firshein 1
PMCID: PMC205740  PMID: 1729239

Abstract

A Bacillus subtilis membrane-associated protein that binds specifically to the origin region of DNA replication may act as an inhibitor of DNA replication (J. Laffan and W. Firshein, Proc. Natl. Acad. Sci. USA 85:7452-7456, 1988). This protein, originally estimated to be 64 kDa, had a slightly lower molecular size (57 kDa), as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis during these studies. The size difference may be due to processing that results in modification of the protein. The protein can be extracted from both cytosol and membrane fractions, and the amounts in these fractions vary during the developmental cycle of B. subtilis. A complex pattern of expression in which significant levels were detected in spores was revealed; levels decreased dramatically during germination and increased after the first round of DNA replication. The decrease during germination was due to protease activity, as demonstrated by the addition of protease inhibitors and radioactive-labeling chase experiments. During vegetative growth, the protein levels increased until stationary phase, after which there was another decrease during sporulation. The decrease during sporulation may be partially due to sequestering of the protein into forespores, since as the putative repressor protein decreased in the mother cell, it increased in the forespores. However, protease activity was also involved in the decrease in the mother cell. The changes in expression of this protein are consistent with its role as a repressor of initiation of DNA replication. Additional studies, including sequence analysis and further antibody analysis, show that this protein is not a subunit of the pyruvate dehydrogenase complex. This relationship had been a possibility based upon the results of others (H. Hemila, A. Pavla, L. Paulin, S. Arvidson, and I. Palva, J. Bacteriol. 172:5052-5063, 1990).

Full text

PDF
477

Images in this article

479Image
on p.479
479Image
on p.479
480Image
on p.480
481Image
on p.481
481Image
on p.481
482Image
on p.482

Selected References

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

  1. Dancer B. N., Mandelstam J. Production and possible function of serine protease during sporulation of Bacillus subtilis. J Bacteriol. 1975 Feb;121(2):406–410. doi: 10.1128/jb.121.2.406-410.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Firshein W. Role of the DNA/membrane complex in prokaryotic DNA replication. Annu Rev Microbiol. 1989;43:89–120. doi: 10.1146/annurev.mi.43.100189.000513. [DOI] [PubMed] [Google Scholar]
  3. Garrick-Silversmith L., Torriani A. Macromolecular syntheses during germination and outgrowth of Bacillus subtilis spores. J Bacteriol. 1973 May;114(2):507–516. doi: 10.1128/jb.114.2.507-516.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hemilä H., Palva A., Paulin L., Arvidson S., Palva I. Secretory S complex of Bacillus subtilis: sequence analysis and identity to pyruvate dehydrogenase. J Bacteriol. 1990 Sep;172(9):5052–5063. doi: 10.1128/jb.172.9.5052-5063.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Horiuchi S., Marty-Mazars D., Tai P. C., Davis B. D. Localization and quantitation of proteins characteristic of the complexed membrane of Bacillus subtilis. J Bacteriol. 1983 Jun;154(3):1215–1221. doi: 10.1128/jb.154.3.1215-1221.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kawamura F., Doi R. H. Construction of a Bacillus subtilis double mutant deficient in extracellular alkaline and neutral proteases. J Bacteriol. 1984 Oct;160(1):442–444. doi: 10.1128/jb.160.1.442-444.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kornberg A., Spudich J. A., Nelson D. L., Deutscher M. P. Origin of proteins in sporulation. Annu Rev Biochem. 1968;37:51–78. doi: 10.1146/annurev.bi.37.070168.000411. [DOI] [PubMed] [Google Scholar]
  8. Köhler E., Antranikian G. Covalent modification of proteins in Bacillus subtilis during the process of sporulation, germination and outgrowth. FEMS Microbiol Lett. 1989 Jan 1;48(1):87–92. doi: 10.1016/0378-1097(89)90152-3. [DOI] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Laffan J. J., Firshein W. Origin-specific DNA-binding membrane-associated protein may be involved in repression of initiation of DNA replication in Bacillus subtilis. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7452–7456. doi: 10.1073/pnas.85.20.7452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mark D. F., Richardson C. C. Escherichia coli thioredoxin: a subunit of bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1976 Mar;73(3):780–784. doi: 10.1073/pnas.73.3.780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Marty-Mazars D., Horiuchi S., Tai P. C., Davis B. D. Proteins of ribosome-bearing and free-membrane domains in Bacillus subtilis. J Bacteriol. 1983 Jun;154(3):1381–1388. doi: 10.1128/jb.154.3.1381-1388.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. OISHI M., YOSHIKAWA H., SUEOKA N. SYNCHRONOUS AND DICHOTOMOUS REPLICATIONS OF THE BACILLUS SUBTILIS CHROMOSOME DURING SPORE GERMINATION. Nature. 1964 Dec 12;204:1069–1073. doi: 10.1038/2041069a0. [DOI] [PubMed] [Google Scholar]
  14. Panzer S., Losick R., Sun D., Setlow P. Evidence for an additional temporal class of gene expression in the forespore compartment of sporulating Bacillus subtilis. J Bacteriol. 1989 Jan;171(1):561–564. doi: 10.1128/jb.171.1.561-564.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Spizizen J. TRANSFORMATION OF BIOCHEMICALLY DEFICIENT STRAINS OF BACILLUS SUBTILIS BY DEOXYRIBONUCLEATE. Proc Natl Acad Sci U S A. 1958 Oct 15;44(10):1072–1078. doi: 10.1073/pnas.44.10.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Spudich J. A., Kornberg A. Biochemical studies of bacterial sporulation and germaination. VII. Protein turnover during sporulation of Bacillus subtilis. J Biol Chem. 1968 Sep 10;243(17):4600–4605. [PubMed] [Google Scholar]
  17. Srere P. A. Complexes of sequential metabolic enzymes. Annu Rev Biochem. 1987;56:89–124. doi: 10.1146/annurev.bi.56.070187.000513. [DOI] [PubMed] [Google Scholar]
  18. Vasantha N., Freese E. Enzyme changes during Bacillus subtilis sporulation caused by deprivation of guanine nucleotides. J Bacteriol. 1980 Dec;144(3):1119–1125. doi: 10.1128/jb.144.3.1119-1125.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Vold B. S. Preparation of tRNA's and aminoacyl-tRNA synthetases from Bacillus subtilis cells at various stages of growth and spores. Methods Enzymol. 1974;29:502–510. doi: 10.1016/0076-6879(74)29045-1. [DOI] [PubMed] [Google Scholar]
  20. YOSHIKAWA H., O'SULLIVAN A., SUEOKA N. SEQUENTIAL REPLICATION OF THE BACILLUS SUBTILIS CHROMOSOME. 3. REGULATION OF INITIATION. Proc Natl Acad Sci U S A. 1964 Oct;52:973–980. doi: 10.1073/pnas.52.4.973. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES