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. 1983 Jun;154(3):1088–1097. doi: 10.1128/jb.154.3.1088-1097.1983

Genetics of Bacillus subtilis chemotaxis: isolation and mapping of mutations and cloning of chemotaxis genes.

G W Ordal, D O Nettleton, J A Hoch
PMCID: PMC217578  PMID: 6222032

Abstract

We isolated a collection of chemotaxis mutants and characterized them for chemotactic phenotype and genotype. The mutations of most of these mutants mapped in the region between pyrD and thyA. However, the mutation in the gene specifying the chemotactic methyltransferase mapped very close to aroF. From a bank of phages containing Bacillus subtilis DNA we identified two lambda charon4 phages that contained genes specifying chemotactic functions. The inserted DNAs were removed by digestion with restriction endonuclease EcoRI and were found to share a 4.0-kilobase (kb) fragment. One of these DNAs also contained a 7.7-kb fragment, and the other also contained a 10.9-kb fragment. We identified mutants that were complemented by each fragment. The fragments were each ligated into plasmid pFH7 and were incorporated into lysogenic SP beta c2 or a deletion mutant of SP beta c2 in order to form transducing phages. The mutants in the collection containing mutations that mapped in the region between pyrD and thyA were tested for complementation by transducing phages containing the 4.0-kb fragment, the 7.7-kb fragment, the 4.0-kb fragment plus the 7.7-kb fragment, and the 10.9-kb fragment. A total of 24 mutants were complemented by the 4.0-kb fragment, 7 were complemented by the 7.7-kb fragment, 9 were complemented by the 4.0-kb fragment plus the 7.7-kb fragment, 15 were complemented by the 10.9-kb fragment, and 25 were complemented by none of the fragments.

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Selected References

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

  1. Adler J. Chemoreceptors in bacteria. Science. 1969 Dec 26;166(3913):1588–1597. doi: 10.1126/science.166.3913.1588. [DOI] [PubMed] [Google Scholar]
  2. Aswad D., Koshland D. E., Jr Isolation, characterization and complementation of Salmonella typhimurium chemotaxis mutants. J Mol Biol. 1975 Sep 15;97(2):225–235. doi: 10.1016/s0022-2836(75)80036-2. [DOI] [PubMed] [Google Scholar]
  3. Berg H. C., Tedesco P. M. Transient response to chemotactic stimuli in Escherichia coli. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3235–3239. doi: 10.1073/pnas.72.8.3235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blattner F. R., Williams B. G., Blechl A. E., Denniston-Thompson K., Faber H. E., Furlong L., Grunwald D. J., Kiefer D. O., Moore D. D., Schumm J. W. Charon phages: safer derivatives of bacteriophage lambda for DNA cloning. Science. 1977 Apr 8;196(4286):161–169. doi: 10.1126/science.847462. [DOI] [PubMed] [Google Scholar]
  5. Boyd A., Simon M. Bacterial chemotaxis. Annu Rev Physiol. 1982;44:501–517. doi: 10.1146/annurev.ph.44.030182.002441. [DOI] [PubMed] [Google Scholar]
  6. Burgess-Cassler A., Ullah A. H., Ordal G. W. Purification and characterization of Bacillus subtilis methyl-accepting chemotaxis protein methyltransferase II. J Biol Chem. 1982 Jul 25;257(14):8412–8417. [PubMed] [Google Scholar]
  7. Clarke S., Koshland D. E., Jr Membrane receptors for aspartate and serine in bacterial chemotaxis. J Biol Chem. 1979 Oct 10;254(19):9695–9702. [PubMed] [Google Scholar]
  8. Dedonder R. A., Lepesant J. A., Lepesant-Kejzlarová J., Billault A., Steinmetz M., Kunst F. Construction of a kit of reference strains for rapid genetic mapping in Bacillus subtilis 168. Appl Environ Microbiol. 1977 Apr;33(4):989–993. doi: 10.1128/aem.33.4.989-993.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ehrlich S. D. DNA cloning in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1433–1436. doi: 10.1073/pnas.75.3.1433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ferrari E., Henner D. J., Hoch J. A. Isolation of Bacillus subtilis genes from a charon 4A library. J Bacteriol. 1981 Apr;146(1):430–432. doi: 10.1128/jb.146.1.430-432.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Frankel R. W., Joys T. M. Adsorption Specificity of Bacteriophage PBS1. J Bacteriol. 1966 Aug;92(2):388–389. doi: 10.1128/jb.92.2.388-389.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Goldman D. J., Ordal G. W. Sensory adaptation and deadaptation by Bacillus subtilis. J Bacteriol. 1981 Jul;147(1):267–270. doi: 10.1128/jb.147.1.267-270.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goldman D. J., Worobec S. W., Siegel R. B., Hecker R. V., Ordal G. W. Chemotaxis in Bacillus subtilis: effects of attractants on the level of methylation of methyl-accepting chemotaxis proteins and the role of demethylation in the adaptation process. Biochemistry. 1982 Mar 2;21(5):915–920. doi: 10.1021/bi00534a016. [DOI] [PubMed] [Google Scholar]
  14. Hedblom M. L., Adler J. Genetic and biochemical properties of Escherichia coli mutants with defects in serine chemotaxis. J Bacteriol. 1980 Dec;144(3):1048–1060. doi: 10.1128/jb.144.3.1048-1060.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Henner D. J., Hoch J. A. The Bacillus subtilis chromosome. Microbiol Rev. 1980 Mar;44(1):57–82. doi: 10.1128/mr.44.1.57-82.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  17. Kemper J. Gene order and co-transduction in the leu-ara-fol-pyrA region of the Salmonella typhimurium linkage map. J Bacteriol. 1974 Jan;117(1):94–99. doi: 10.1128/jb.117.1.94-99.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kleene S. J., Toews M. L., Adler J. Isolation of glutamic acid methyl ester from an Escherichia coli membrane protein involved in chemotaxis. J Biol Chem. 1977 May 25;252(10):3214–3218. [PubMed] [Google Scholar]
  19. Kort E. N., Goy M. F., Larsen S. H., Adler J. Methylation of a membrane protein involved in bacterial chemotaxis. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3939–3943. doi: 10.1073/pnas.72.10.3939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Macnab R. M., Koshland D. E., Jr The gradient-sensing mechanism in bacterial chemotaxis. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2509–2512. doi: 10.1073/pnas.69.9.2509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ordal G. W., Adler J. Isolation and complementation of mutants in galactose taxis and transport. J Bacteriol. 1974 Feb;117(2):509–516. doi: 10.1128/jb.117.2.509-516.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ordal G. W. Effect of methionine on chemotaxis by Bacillus subtilis. J Bacteriol. 1976 Mar;125(3):1005–1012. doi: 10.1128/jb.125.3.1005-1012.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ordal G. W., Goldman D. J. Chemotactic repellents of Bacillus subtilis. J Mol Biol. 1976 Jan 5;100(1):103–108. doi: 10.1016/s0022-2836(76)80037-x. [DOI] [PubMed] [Google Scholar]
  24. Ordal G. W., Goldman D. J. Chemotaxis away from uncouplers of oxidative phosphorylation in Bacillus subtilis. Science. 1975 Sep 5;189(4205):802–805. doi: 10.1126/science.808854. [DOI] [PubMed] [Google Scholar]
  25. Ordal G. W. Recognition sites for chemotactic repellents of Bacillus subtilis. J Bacteriol. 1976 Apr;126(1):72–79. doi: 10.1128/jb.126.1.72-79.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ordal G. W., Villani D. P., Gibson K. J. Amino acid chemoreceptors of Bacillus subtilis. J Bacteriol. 1977 Jan;129(1):156–165. doi: 10.1128/jb.129.1.156-165.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Parkinson J. S. Behavioral genetics in bacteria. Annu Rev Genet. 1977;11:397–414. doi: 10.1146/annurev.ge.11.120177.002145. [DOI] [PubMed] [Google Scholar]
  28. Parkinson J. S., Huskey R. J. Deletion mutants of bacteriophage lambda. I. Isolation and initial characterization. J Mol Biol. 1971 Mar 14;56(2):369–384. doi: 10.1016/0022-2836(71)90471-2. [DOI] [PubMed] [Google Scholar]
  29. Parkinson J. S., Revello P. T. Sensory adaptation mutants of E. coli. Cell. 1978 Dec;15(4):1221–1230. doi: 10.1016/0092-8674(78)90048-x. [DOI] [PubMed] [Google Scholar]
  30. Rosenthal R., Toye P. A., Korman R. Z., Zahler S. A. The prophage of SP beta c2dcitK1, A defective specialized transducing phage of Bacillus subtilis. Genetics. 1979 Jul;92(3):721–739. doi: 10.1093/genetics/92.3.721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Silverman M., Simon M. Chemotaxis in Escherichia coli: methylation of che gene products. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3317–3321. doi: 10.1073/pnas.74.8.3317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Springer M. S., Goy M. F., Adler J. Sensory transduction in Escherichia coli: two complementary pathways of information processing that involve methylated proteins. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3312–3316. doi: 10.1073/pnas.74.8.3312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Taylor B. L., Miller J. B., Warrick H. M., Koshland D. E., Jr Electron acceptor taxis and blue light effect on bacterial chemotaxis. J Bacteriol. 1979 Nov;140(2):567–573. doi: 10.1128/jb.140.2.567-573.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tsang N., Macnab R., Koshland D. E., Jr Common mechanism for repellents and attractants in bacterial chemotaxis. Science. 1973 Jul 6;181(4094):60–63. doi: 10.1126/science.181.4094.60. [DOI] [PubMed] [Google Scholar]
  36. Ullah A. H., Ordal G. W. In vivo and in vitro chemotactic methylation in Bacillus subtilis. J Bacteriol. 1981 Feb;145(2):958–965. doi: 10.1128/jb.145.2.958-965.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Van Der Werf P., Koshland D. E., Jr Identification of a gamma-glutamyl methyl ester in bacterial membrane protein involved in chemotaxis. J Biol Chem. 1977 Apr 25;252(8):2793–2795. [PubMed] [Google Scholar]
  38. Zahler S. A., Korman R. Z., Rosenthal R., Hemphill H. E. Bacillus subtilis bacteriophage SPbeta: localization of the prophage attachment site, and specialized transduction. J Bacteriol. 1977 Jan;129(1):556–558. doi: 10.1128/jb.129.1.556-558.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]

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