Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Jul;81(14):4490–4494. doi: 10.1073/pnas.81.14.4490

Cell-division control in Escherichia coli: specific induction of the SOS function SfiA protein is sufficient to block septation.

O Huisman, R D'Ari, S Gottesman
PMCID: PMC345616  PMID: 6087326

Abstract

Blocks in DNA replication cause a rapid arrest of cell division in Escherichia coli. We have previously established that the function SfiA (SulA), induced under these conditions as part of the SOS response, is involved in this inhibition of division. To separate the effects of SfiA from those of other SOS functions, we have constructed a plac-sfiA operon fusion, permitting specific induction of SfiA protein by addition of the lac operon inducer isopropyl beta-D-thiogalactopyranoside (IPTG). In lon mutants, in which the unstable SfiA protein has a longer half-life, IPTG caused a rapid arrest of cell division. Under these conditions, there is no concomitant induction of the SOS response. IPTG also caused a rapid arrest of cell division in lon+ strains. These results demonstrate that induction of the SfiA protein is sufficient to cause inhibition of division. Mutations in the sfiB gene can suppress IPTG-induced SfiA-mediated inhibition of division. At higher SfiA concentrations, however, even sfiB mutants cease division; an additional mutation genetically inseparable from sfiB restores normal division. These observations reinforce the hypothesis that the SfiB protein, probably required for cell septation, is the target of action of the SfiA division inhibitor.

Full text

PDF
4491

Images in this article

Selected References

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

  1. Beck E., Bremer E. Nucleotide sequence of the gene ompA coding the outer membrane protein II of Escherichia coli K-12. Nucleic Acids Res. 1980 Jul 11;8(13):3011–3027. doi: 10.1093/nar/8.13.3011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bremer E., Beck E., Hindennach I., Sonntag I., Henning U. Cloned structural gene (ompA) for an integral outer membrane protein of Escherichia coli K-12: localization on hybrid plasmid pTU100 and expression of a fragment of the gene. Mol Gen Genet. 1980;179(1):13–20. doi: 10.1007/BF00268440. [DOI] [PubMed] [Google Scholar]
  3. Burton P., Holland I. B. Two pathways of division inhibition in UV-irradiated E. coli. Mol Gen Genet. 1983;190(1):128–132. doi: 10.1007/BF00330334. [DOI] [PubMed] [Google Scholar]
  4. Charette M. F., Henderson G. W., Markovitz A. ATP hydrolysis-dependent protease activity of the lon (capR) protein of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4728–4732. doi: 10.1073/pnas.78.8.4728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chung C. H., Goldberg A. L. The product of the lon (capR) gene in Escherichia coli is the ATP-dependent protease, protease La. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4931–4935. doi: 10.1073/pnas.78.8.4931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen S. S., Barner H. D. STUDIES ON UNBALANCED GROWTH IN ESCHERICHIA COLI. Proc Natl Acad Sci U S A. 1954 Oct;40(10):885–893. doi: 10.1073/pnas.40.10.885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. D'Ari R., Huisman O. Novel mechanism of cell division inhibition associated with the SOS response in Escherichia coli. J Bacteriol. 1983 Oct;156(1):243–250. doi: 10.1128/jb.156.1.243-250.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gayda R. C., Yamamoto L. T., Markovitz A. Second-site mutations in capR (lon) strains of Escherichia coli K-12 that prevent radiation sensitivity and allow bacteriophage lambda to lysogenize. J Bacteriol. 1976 Sep;127(3):1208–1216. doi: 10.1128/jb.127.3.1208-1216.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. George J., Castellazzi M., Buttin G. Prophage induction and cell division in E. coli. III. Mutations sfiA and sfiB restore division in tif and lon strains and permit the expression of mutator properties of tif. Mol Gen Genet. 1975 Oct 22;140(4):309–332. [PubMed] [Google Scholar]
  10. Gottesman S., Gottesman M., Shaw J. E., Pearson M. L. Protein degradation in E. coli: the lon mutation and bacteriophage lambda N and cII protein stability. Cell. 1981 Apr;24(1):225–233. doi: 10.1016/0092-8674(81)90518-3. [DOI] [PubMed] [Google Scholar]
  11. Gottesman S., Halpern E., Trisler P. Role of sulA and sulB in filamentation by lon mutants of Escherichia coli K-12. J Bacteriol. 1981 Oct;148(1):265–273. doi: 10.1128/jb.148.1.265-273.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gottesman S., Zipser D. Deg phenotype of Escherichia coli lon mutants. J Bacteriol. 1978 Feb;133(2):844–851. doi: 10.1128/jb.133.2.844-851.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hirota Y., Ryter A., Jacob F. Thermosensitive mutants of E. coli affected in the processes of DNA synthesis and cellular division. Cold Spring Harb Symp Quant Biol. 1968;33:677–693. doi: 10.1101/sqb.1968.033.01.077. [DOI] [PubMed] [Google Scholar]
  14. Huisman O., D'Ari R. An inducible DNA replication-cell division coupling mechanism in E. coli. Nature. 1981 Apr 30;290(5809):797–799. doi: 10.1038/290797a0. [DOI] [PubMed] [Google Scholar]
  15. Huisman O., D'Ari R. Effect of suppressors of SOS-mediated filamentation on sfiA operon expression in Escherichia coli. J Bacteriol. 1983 Jan;153(1):169–175. doi: 10.1128/jb.153.1.169-175.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Huisman O., D'Ari R., George J. Further characterization of sfiA and sfiB mutations in Escherichia coli. J Bacteriol. 1980 Oct;144(1):185–191. doi: 10.1128/jb.144.1.185-191.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Huisman O., D'Ari R., George J. Inducible sfi dependent division inhibition in Escherichia coli. Mol Gen Genet. 1980;177(4):629–636. doi: 10.1007/BF00272673. [DOI] [PubMed] [Google Scholar]
  18. Huisman O., Jacques M., D'ari R., Caro L. Role of the sfiA-dependent cell division regulation system in Escherichia coli. J Bacteriol. 1983 Feb;153(2):1072–1074. doi: 10.1128/jb.153.2.1072-1074.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Johnson B. F. Fine structure mapping and properties of mutations suppressing the lon mutation in Escherichia coli K-12 and B strains. Genet Res. 1977 Dec;30(3):273–286. doi: 10.1017/s0016672300017687. [DOI] [PubMed] [Google Scholar]
  20. Little J. W., Edmiston S. H., Pacelli L. Z., Mount D. W. Cleavage of the Escherichia coli lexA protein by the recA protease. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3225–3229. doi: 10.1073/pnas.77.6.3225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Little J. W., Mount D. W. The SOS regulatory system of Escherichia coli. Cell. 1982 May;29(1):11–22. doi: 10.1016/0092-8674(82)90085-x. [DOI] [PubMed] [Google Scholar]
  22. Lutkenhaus J. F. Coupling of DNA replication and cell division: sulB is an allele of ftsZ. J Bacteriol. 1983 Jun;154(3):1339–1346. doi: 10.1128/jb.154.3.1339-1346.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mackie G., Wilson D. B. Regulation of the gal operon of Escherichia coli by the capR gene. J Biol Chem. 1972 May 25;247(10):2973–2978. [PubMed] [Google Scholar]
  24. Mizusawa S., Court D., Gottesman S. Transcription of the sulA gene and repression by LexA. J Mol Biol. 1983 Dec 15;171(3):337–343. doi: 10.1016/0022-2836(83)90097-9. [DOI] [PubMed] [Google Scholar]
  25. Mizusawa S., Gottesman S. Protein degradation in Escherichia coli: the lon gene controls the stability of sulA protein. Proc Natl Acad Sci U S A. 1983 Jan;80(2):358–362. doi: 10.1073/pnas.80.2.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mizusawa S., Ward D. F. A bacteriophage lambda vector for cloning with BamHI and Sau3A. Gene. 1982 Dec;20(3):317–322. doi: 10.1016/0378-1119(82)90200-1. [DOI] [PubMed] [Google Scholar]
  27. Shineberg B., Zipser D. The ion gene and degradation of beta-galactosidase nonsense fragments. J Bacteriol. 1973 Dec;116(3):1469–1471. doi: 10.1128/jb.116.3.1469-1471.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
  29. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  30. Walker J. R., Kovarik A., Allen J. S., Gustafson R. A. Regulation of bacterial cell division: temperature-sensitive mutants of Escherichia coli that are defective in septum formation. J Bacteriol. 1975 Aug;123(2):693–703. doi: 10.1128/jb.123.2.693-703.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Witkin E. M. The radiation sensitivity of Escherichia coli B: a hypothesis relating filament formation and prophage induction. Proc Natl Acad Sci U S A. 1967 May;57(5):1275–1279. doi: 10.1073/pnas.57.5.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Witkin E. M. Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev. 1976 Dec;40(4):869–907. doi: 10.1128/br.40.4.869-907.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES