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. 1992 Dec;174(23):7517–7526. doi: 10.1128/jb.174.23.7517-7526.1992

Identification and characterization of the smbA gene, a suppressor of the mukB null mutant of Escherichia coli.

K Yamanaka 1, T Ogura 1, H Niki 1, S Hiraga 1
PMCID: PMC207461  PMID: 1447125

Abstract

The mukB gene encodes a protein involved in chromosome partitioning in Escherichia coli. To study the function of this protein, we isolated from the temperature-sensitive mukB null mutant and characterized 56 suppressor mutants which could grow at 42 degrees C. Ten of the mutants also showed cold-sensitive growth at 22 degrees C. Using one of the cold-sensitive mutants as host, the wild type of the suppressor gene was cloned. The cloned suppressor gene complemented all of the 56 suppressor mutations. DNA sequencing revealed the presence of an open reading frame of 723 bp which could encode a protein of 25,953 Da. The gene product was indeed detected. The previously undiscovered gene, named smbA (suppressor of mukB), is located at 4 min on the E. coli chromosome, between the tsf and frr genes. The smbA gene is essential for cell proliferation in the range from 22 to 42 degrees C. Cells which lacked the SmbA protein ceased macromolecular synthesis. The smbA mutants are sensitive to a detergent, sodium dodecyl sulfate, and they show a novel morphological phenotype under nonpermissive conditions, suggesting a defect in specific membrane sites.

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Selected References

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

  1. Aldea M., Hernández-Chico C., de la Campa A. G., Kushner S. R., Vicente M. Identification, cloning, and expression of bolA, an ftsZ-dependent morphogene of Escherichia coli. J Bacteriol. 1988 Nov;170(11):5169–5176. doi: 10.1128/jb.170.11.5169-5176.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. An G., Bendiak D. S., Mamelak L. A., Friesen J. D. Organization and nucleotide sequence of a new ribosomal operon in Escherichia coli containing the genes for ribosomal protein S2 and elongation factor Ts. Nucleic Acids Res. 1981 Aug 25;9(16):4163–4172. doi: 10.1093/nar/9.16.4163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baird L., Lipinska B., Raina S., Georgopoulos C. Identification of the Escherichia coli sohB gene, a multicopy suppressor of the HtrA (DegP) null phenotype. J Bacteriol. 1991 Sep;173(18):5763–5770. doi: 10.1128/jb.173.18.5763-5770.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Begg K. J., Donachie W. D. Experiments on chromosome separation and positioning in Escherichia coli. New Biol. 1991 May;3(5):475–486. [PubMed] [Google Scholar]
  5. Ben-Bassat A., Bauer K., Chang S. Y., Myambo K., Boosman A., Chang S. Processing of the initiation methionine from proteins: properties of the Escherichia coli methionine aminopeptidase and its gene structure. J Bacteriol. 1987 Feb;169(2):751–757. doi: 10.1128/jb.169.2.751-757.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Blakely G., Colloms S., May G., Burke M., Sherratt D. Escherichia coli XerC recombinase is required for chromosomal segregation at cell division. New Biol. 1991 Aug;3(8):789–798. [PubMed] [Google Scholar]
  8. Brady G., Jantzen H. M., Bernard H. U., Brown R., Schütz G., Hashimoto-Gotoh T. New cosmid vectors developed for eukaryotic DNA cloning. Gene. 1984 Feb;27(2):223–232. doi: 10.1016/0378-1119(84)90143-4. [DOI] [PubMed] [Google Scholar]
  9. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Clerget M. Site-specific recombination promoted by a short DNA segment of plasmid R1 and by a homologous segment in the terminus region of the Escherichia coli chromosome. New Biol. 1991 Aug;3(8):780–788. [PubMed] [Google Scholar]
  11. Cook W. R., Kepes F., Joseleau-Petit D., MacAlister T. J., Rothfield L. I. Proposed mechanism for generation and localization of new cell division sites during the division cycle of Escherichia coli. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7144–7148. doi: 10.1073/pnas.84.20.7144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Donachie W. D., Begg K. J. Chromosome partition in Escherichia coli requires postreplication protein synthesis. J Bacteriol. 1989 Oct;171(10):5405–5409. doi: 10.1128/jb.171.10.5405-5409.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hashimoto-Gotoh T., Franklin F. C., Nordheim A., Timmis K. N. Specific-purpose plasmid cloning vectors. I. Low copy number, temperature-sensitive, mobilization-defective pSC101-derived containment vectors. Gene. 1981 Dec;16(1-3):227–235. doi: 10.1016/0378-1119(81)90079-2. [DOI] [PubMed] [Google Scholar]
  14. Hiraga S., Niki H., Ogura T., Ichinose C., Mori H., Ezaki B., Jaffé A. Chromosome partitioning in Escherichia coli: novel mutants producing anucleate cells. J Bacteriol. 1989 Mar;171(3):1496–1505. doi: 10.1128/jb.171.3.1496-1505.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hiraga S., Ogura T., Niki H., Ichinose C., Mori H. Positioning of replicated chromosomes in Escherichia coli. J Bacteriol. 1990 Jan;172(1):31–39. doi: 10.1128/jb.172.1.31-39.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Hussain K., Begg K. J., Salmond G. P., Donachie W. D. ParD: a new gene coding for a protein required for chromosome partitioning and septum localization in Escherichia coli. Mol Microbiol. 1987 Jul;1(1):73–81. doi: 10.1111/j.1365-2958.1987.tb00529.x. [DOI] [PubMed] [Google Scholar]
  18. Hussain K., Elliott E. J., Salmond G. P. The parD- mutant of Escherichia coli also carries a gyrAam mutation. The complete sequence of gyrA. Mol Microbiol. 1987 Nov;1(3):259–273. doi: 10.1111/j.1365-2958.1987.tb01932.x. [DOI] [PubMed] [Google Scholar]
  19. Ichikawa S., Kaji A. Molecular cloning and expression of ribosome releasing factor. J Biol Chem. 1989 Nov 25;264(33):20054–20059. [PubMed] [Google Scholar]
  20. Ichikawa S., Ryoji M., Siegfried Z., Kaji A. Localization of the ribosome-releasing factor gene in the Escherichia coli chromosome. J Bacteriol. 1989 Jul;171(7):3689–3695. doi: 10.1128/jb.171.7.3689-3695.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kang P. J., Craig E. A. Identification and characterization of a new Escherichia coli gene that is a dosage-dependent suppressor of a dnaK deletion mutation. J Bacteriol. 1990 Apr;172(4):2055–2064. doi: 10.1128/jb.172.4.2055-2064.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kato J., Nishimura Y., Imamura R., Niki H., Hiraga S., Suzuki H. New topoisomerase essential for chromosome segregation in E. coli. Cell. 1990 Oct 19;63(2):393–404. doi: 10.1016/0092-8674(90)90172-b. [DOI] [PubMed] [Google Scholar]
  23. Kato J., Nishimura Y., Suzuki H. Escherichia coli parA is an allele of the gyrB gene. Mol Gen Genet. 1989 May;217(1):178–181. doi: 10.1007/BF00330959. [DOI] [PubMed] [Google Scholar]
  24. Kato J., Nishimura Y., Yamada M., Suzuki H., Hirota Y. Gene organization in the region containing a new gene involved in chromosome partition in Escherichia coli. J Bacteriol. 1988 Sep;170(9):3967–3977. doi: 10.1128/jb.170.9.3967-3977.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  26. Kuempel P. L., Henson J. M., Dircks L., Tecklenburg M., Lim D. F. dif, a recA-independent recombination site in the terminus region of the chromosome of Escherichia coli. New Biol. 1991 Aug;3(8):799–811. [PubMed] [Google Scholar]
  27. Kusukawa N., Yura T. Heat shock protein GroE of Escherichia coli: key protective roles against thermal stress. Genes Dev. 1988 Jul;2(7):874–882. doi: 10.1101/gad.2.7.874. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Louarn J. M., Louarn J., François V., Patte J. Analysis and possible role of hyperrecombination in the termination region of the Escherichia coli chromosome. J Bacteriol. 1991 Aug;173(16):5097–5104. doi: 10.1128/jb.173.16.5097-5104.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Morona R., Reeves P. The tolC locus of Escherichia coli affects the expression of three major outer membrane proteins. J Bacteriol. 1982 Jun;150(3):1016–1023. doi: 10.1128/jb.150.3.1016-1023.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Niki H., Jaffé A., Imamura R., Ogura T., Hiraga S. The new gene mukB codes for a 177 kd protein with coiled-coil domains involved in chromosome partitioning of E. coli. EMBO J. 1991 Jan;10(1):183–193. doi: 10.1002/j.1460-2075.1991.tb07935.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Niki H., Ogura T., Hiraga S. Linear multimer formation of plasmid DNA in Escherichia coli hopE (recD) mutants. Mol Gen Genet. 1990 Oct;224(1):1–9. doi: 10.1007/BF00259444. [DOI] [PubMed] [Google Scholar]
  33. Obar R. A., Collins C. A., Hammarback J. A., Shpetner H. S., Vallee R. B. Molecular cloning of the microtubule-associated mechanochemical enzyme dynamin reveals homology with a new family of GTP-binding proteins. Nature. 1990 Sep 20;347(6290):256–261. doi: 10.1038/347256a0. [DOI] [PubMed] [Google Scholar]
  34. Orr E., Fairweather N. F., Holland I. B., Pritchard R. H. Isolation and characterisation of a strain carrying a conditional lethal mutation in the cou gene of Escherichia coli K12. Mol Gen Genet. 1979;177(1):103–112. doi: 10.1007/BF00267259. [DOI] [PubMed] [Google Scholar]
  35. Pedersen S., Skouv J., Kajitani M., Ishihama A. Transcriptional organization of the rpsA operon of Escherichia coli. Mol Gen Genet. 1984;196(1):135–140. doi: 10.1007/BF00334105. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Sargent M. G. Nuclear segregation in Bacillus subtilis. Nature. 1974 Jul 19;250(463):252–254. doi: 10.1038/250252a0. [DOI] [PubMed] [Google Scholar]
  38. Seelke R., Kline B., Aleff R., Porter R. D., Shields M. S. Mutations in the recD gene of Escherichia coli that raise the copy number of certain plasmids. J Bacteriol. 1987 Oct;169(10):4841–4844. doi: 10.1128/jb.169.10.4841-4844.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shimizu I., Kaji A. Identification of the promoter region of the ribosome-releasing factor cistron (frr). J Bacteriol. 1991 Aug;173(16):5181–5187. doi: 10.1128/jb.173.16.5181-5187.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Steck T. R., Drlica K. Bacterial chromosome segregation: evidence for DNA gyrase involvement in decatenation. Cell. 1984 Apr;36(4):1081–1088. doi: 10.1016/0092-8674(84)90058-8. [DOI] [PubMed] [Google Scholar]
  41. Takeshita S., Sato M., Toba M., Masahashi W., Hashimoto-Gotoh T. High-copy-number and low-copy-number plasmid vectors for lacZ alpha-complementation and chloramphenicol- or kanamycin-resistance selection. Gene. 1987;61(1):63–74. doi: 10.1016/0378-1119(87)90365-9. [DOI] [PubMed] [Google Scholar]
  42. Vicente M., Kushner S. R., Garrido T., Aldea M. The role of the 'gearbox' in the transcription of essential genes. Mol Microbiol. 1991 Sep;5(9):2085–2091. doi: 10.1111/j.1365-2958.1991.tb02137.x. [DOI] [PubMed] [Google Scholar]
  43. Wandersman C., Delepelaire P. TolC, an Escherichia coli outer membrane protein required for hemolysin secretion. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4776–4780. doi: 10.1073/pnas.87.12.4776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Webster R. E. The tol gene products and the import of macromolecules into Escherichia coli. Mol Microbiol. 1991 May;5(5):1005–1011. doi: 10.1111/j.1365-2958.1991.tb01873.x. [DOI] [PubMed] [Google Scholar]
  45. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  46. 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]
  47. 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]

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