Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1981 Feb;145(2):958–965. doi: 10.1128/jb.145.2.958-965.1981

In vivo and in vitro chemotactic methylation in Bacillus subtilis.

A H Ullah, G W Ordal
PMCID: PMC217204  PMID: 6780537

Abstract

Two doublets of Bacillus subtilis membrane proteins with molecular weights of 69,000 and 71,000 and of 30,000 and 30,800, were labeled by C3H3 transfer in the absence of protein synthesis. In addition, there was intense methylation of several low-molecular-weight substances. Both doublets were missing in a chemotaxis mutant. The equivalent proteins in Escherichia coli and Salmonella typhimurium are believed to be the methyl-accepting chemotaxis proteins. The higher-molecular-weight doublet bands were increased in degree of methylation upon addition of attractant to the bacteria. A methyltransferase from B. subtilis that methylates the wild-type membrane significantly better than the mutant membrane, using S-adenosylmethionine, has been partly purified. The methylated product was alkali labile and is probably a gamma-glutamyl methyl ester, as in E. coli and S. typhimurium. Ca2+ ion inhibited the methyltransferase, with a Ki of about 80 nM. Analysis of the in vitro methylation product showed labeling of the 69,000-dalton methyl-accepting chemotaxis protein and a low-molecular-weight protein, using wild-type membrane. Labeling of the low-molecular-weight protein but not of the 69,000 dalton protein was observed when the mutant membrane was used. The chemotaxis mutant tumbled much longer than the wild type when diluted away from attractant.

Full text

PDF
958

Images in this article

960Image
on p.960
960Image
on p.960
961Image
on p.961
961Image
on p.961

Selected References

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

  1. Ames G. F. Lipids of Salmonella typhimurium and Escherichia coli: structure and metabolism. J Bacteriol. 1968 Mar;95(3):833–843. doi: 10.1128/jb.95.3.833-843.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Armstrong J. B. An S-adenosylmethionine requirement for chemotaxis in Escherichia coli. Can J Microbiol. 1972 Nov;18(11):1695–1701. doi: 10.1139/m72-263. [DOI] [PubMed] [Google Scholar]
  3. Armstrong J. B. Chemotaxis and methionine metabolism in Escherichia coli. Can J Microbiol. 1972 May;18(5):591–596. doi: 10.1139/m72-093. [DOI] [PubMed] [Google Scholar]
  4. Aswad D. W., Koshland D. E., Jr Evidence for an S-adenosylmethionine requirement in the chemotactic behavior of Salmonella typhimurium. J Mol Biol. 1975 Sep 15;97(2):207–223. doi: 10.1016/s0022-2836(75)80035-0. [DOI] [PubMed] [Google Scholar]
  5. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  6. Barrett J. C., Crawford B. D., Ts'o P. O. Quantitation of fibrinolytic activity of Syrian hamster fibroblasts using 3H-labeled fibrinogen prepared by reductive alkylation. Cancer Res. 1977 Apr;37(4):1182–1185. [PubMed] [Google Scholar]
  7. Berg H. C., Brown D. A. Chemotaxis in Escherichia coli analysed by three-dimensional tracking. Nature. 1972 Oct 27;239(5374):500–504. doi: 10.1038/239500a0. [DOI] [PubMed] [Google Scholar]
  8. Goy M. F., Springer M. S., Adler J. Sensory transduction in Escherichia coli: role of a protein methylation reaction in sensory adaptation. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4964–4968. doi: 10.1073/pnas.74.11.4964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hazelbauer G. L., Engström P. Parallel pathways for transduction of chemotactic signals in Escherichia coli. Nature. 1980 Jan 3;283(5742):98–100. doi: 10.1038/283098a0. [DOI] [PubMed] [Google Scholar]
  10. Hazelbauer G. L., Mesibov R. E., Adler J. Escherichia coli mutants defective in chemotaxis toward specific chemicals. Proc Natl Acad Sci U S A. 1969 Dec;64(4):1300–1307. doi: 10.1073/pnas.64.4.1300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kim S., Paik W. K. Labile protein-methyl ester: comparison between chemically and enzymatically synthesized. Experientia. 1976 Aug 15;32(8):982–984. doi: 10.1007/BF01933924. [DOI] [PubMed] [Google Scholar]
  12. Kleene S. J., Hobson A. C., Adler J. Attractants and repellents influence methylation and demethylation of methyl-accepting chemotaxis proteins in an extract of Escherichia coli. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6309–6313. doi: 10.1073/pnas.76.12.6309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. 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]
  16. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Mesibov R., Adler J. Chemotaxis toward amino acids in Escherichia coli. J Bacteriol. 1972 Oct;112(1):315–326. doi: 10.1128/jb.112.1.315-326.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nakayama T., Munoz L., Doi R. H. A procedure to remove protease activities from Bacillus subtilis sporulating cells and their crude extracts. Anal Biochem. 1977 Mar;78(1):165–170. doi: 10.1016/0003-2697(77)90020-3. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Ordal G. W. Calcium ion regulates chemotactic behaviour in bacteria. Nature. 1977 Nov 3;270(5632):66–67. doi: 10.1038/270066a0. [DOI] [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., Fields R. B. A biochemical mechanism for bacterial chemotaxis. J Theor Biol. 1977 Oct 21;68(4):491–500. doi: 10.1016/0022-5193(77)90100-x. [DOI] [PubMed] [Google Scholar]
  24. Ordal G. W., Gibson K. J. Chemotaxis toward amino acids by Bacillus subtilis. J Bacteriol. 1977 Jan;129(1):151–155. doi: 10.1128/jb.129.1.151-155.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  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. Ridgway H. G., Silverman M., Simon M. I. Localization of proteins controlling motility and chemotaxis in Escherichia coli. J Bacteriol. 1977 Nov;132(2):657–665. doi: 10.1128/jb.132.2.657-665.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. 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]
  30. Springer W. R., Koshland D. E., Jr Identification of a protein methyltransferase as the cheR gene product in the bacterial sensing system. Proc Natl Acad Sci U S A. 1977 Feb;74(2):533–537. doi: 10.1073/pnas.74.2.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Toews M. L., Goy M. F., Springer M. S., Adler J. Attractants and repellents control demethylation of methylated chemotaxis proteins in Escherichia coli. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5544–5548. doi: 10.1073/pnas.76.11.5544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]

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

RESOURCES