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Proc Natl Acad Sci U S A. 1993 Oct 1; 90(19): 8787–8791.

Phosphorylation-dependent binding of a signal molecule to the flagellar switch of bacteria.


Regulation of the direction of flagellar rotation is central to the mechanism of bacterial chemotaxis. The transitions between counterclockwise and clockwise rotation are controlled by a "switch complex" composed of three proteins (FliG, FliM, and FliN) and located at the base of the flagellar motor. The mechanism of function of the switch is unknown. Here we demonstrate that the diffusible clockwise-signal molecule, the CheY protein, binds to the switch, that the primary docking site is FliM, that the extent of CheY binding to FliM is dependent upon the phosphorylation level of CheY, and that it is unaffected by the other two switch proteins. This study provides a biochemical demonstration of binding of a signal molecule to the bacterial switch and demonstrates directly that phosphorylation regulates the activity of this molecule.

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  • Macnab RM, Koshland DE., Jr The gradient-sensing mechanism in bacterial chemotaxis. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2509–2512. [PMC free article] [PubMed]
  • Berg HC, Brown DA. Chemotaxis in Escherichia coli analysed by three-dimensional tracking. Nature. 1972 Oct 27;239(5374):500–504. [PubMed]
  • Adler J. How motile bacteria are attracted and repelled by chemicals: an approach to neurobiology. Lecture held on the occasion of the receipt of the Otto-Warburg-Medaille 1986. Biol Chem Hoppe Seyler. 1987 Mar;368(3):163–173. [PubMed]
  • Bourret RB, Borkovich KA, Simon MI. Signal transduction pathways involving protein phosphorylation in prokaryotes. Annu Rev Biochem. 1991;60:401–441. [PubMed]
  • Stock JB, Surette MG, McCleary WR, Stock AM. Signal transduction in bacterial chemotaxis. J Biol Chem. 1992 Oct 5;267(28):19753–19756. [PubMed]
  • Borkovich KA, Kaplan N, Hess JF, Simon MI. Transmembrane signal transduction in bacterial chemotaxis involves ligand-dependent activation of phosphate group transfer. Proc Natl Acad Sci U S A. 1989 Feb;86(4):1208–1212. [PMC free article] [PubMed]
  • Ninfa EG, Stock A, Mowbray S, Stock J. Reconstitution of the bacterial chemotaxis signal transduction system from purified components. J Biol Chem. 1991 May 25;266(15):9764–9770. [PubMed]
  • Yamaguchi S, Aizawa S, Kihara M, Isomura M, Jones CJ, Macnab RM. Genetic evidence for a switching and energy-transducing complex in the flagellar motor of Salmonella typhimurium. J Bacteriol. 1986 Dec;168(3):1172–1179. [PMC free article] [PubMed]
  • Barak R, Eisenbach M. Correlation between phosphorylation of the chemotaxis protein CheY and its activity at the flagellar motor. Biochemistry. 1992 Feb 18;31(6):1821–1826. [PubMed]
  • Clegg DO, Koshland DE., Jr The role of a signaling protein in bacterial sensing: behavioral effects of increased gene expression. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5056–5060. [PMC free article] [PubMed]
  • Wolfe AJ, Conley MP, Kramer TJ, Berg HC. Reconstitution of signaling in bacterial chemotaxis. J Bacteriol. 1987 May;169(5):1878–1885. [PMC free article] [PubMed]
  • Smith JM, Rowsell EH, Shioi J, Taylor BL. Identification of a site of ATP requirement for signal processing in bacterial chemotaxis. J Bacteriol. 1988 Jun;170(6):2698–2704. [PMC free article] [PubMed]
  • Ravid S, Matsumura P, Eisenbach M. Restoration of flagellar clockwise rotation in bacterial envelopes by insertion of the chemotaxis protein CheY. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7157–7161. [PMC free article] [PubMed]
  • Parkinson JS, Parker SR, Talbert PB, Houts SE. Interactions between chemotaxis genes and flagellar genes in Escherichia coli. J Bacteriol. 1983 Jul;155(1):265–274. [PMC free article] [PubMed]
  • Roman SJ, Meyers M, Volz K, Matsumura P. A chemotactic signaling surface on CheY defined by suppressors of flagellar switch mutations. J Bacteriol. 1992 Oct;174(19):6247–6255. [PMC free article] [PubMed]
  • Sockett H, Yamaguchi S, Kihara M, Irikura VM, Macnab RM. Molecular analysis of the flagellar switch protein FliM of Salmonella typhimurium. J Bacteriol. 1992 Feb;174(3):793–806. [PMC free article] [PubMed]
  • Lukat GS, Lee BH, Mottonen JM, Stock AM, Stock JB. Roles of the highly conserved aspartate and lysine residues in the response regulator of bacterial chemotaxis. J Biol Chem. 1991 May 5;266(13):8348–8354. [PubMed]
  • Barak R, Welch M, Yanovsky A, Oosawa K, Eisenbach M. Acetyladenylate or its derivative acetylates the chemotaxis protein CheY in vitro and increases its activity at the flagellar switch. Biochemistry. 1992 Oct 20;31(41):10099–10107. [PubMed]
  • Bourret RB, Hess JF, Simon MI. Conserved aspartate residues and phosphorylation in signal transduction by the chemotaxis protein CheY. Proc Natl Acad Sci U S A. 1990 Jan;87(1):41–45. [PMC free article] [PubMed]
  • Studier FW, Rosenberg AH, Dunn JJ, Dubendorff JW. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. [PubMed]
  • Tabor S, Richardson CC. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. [PMC free article] [PubMed]
  • KAISER AD, HOGNESS DS. The transformation of Escherichia coli with deoxyribonucleic acid isolated from bacteriophage lambda-dg. J Mol Biol. 1960 Dec;2:392–415. [PubMed]
  • Lukat GS, McCleary WR, Stock AM, Stock JB. Phosphorylation of bacterial response regulator proteins by low molecular weight phospho-donors. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):718–722. [PMC free article] [PubMed]
  • Marwan W, Schäfer W, Oesterhelt D. Signal transduction in Halobacterium depends on fumarate. EMBO J. 1990 Feb;9(2):355–362. [PMC free article] [PubMed]
  • Barak R, Eisenbach M. Fumarate or a fumarate metabolite restores switching ability to rotating flagella of bacterial envelopes. J Bacteriol. 1992 Jan;174(2):643–645. [PMC free article] [PubMed]
  • Lukat GS, Stock AM, Stock JB. Divalent metal ion binding to the CheY protein and its significance to phosphotransfer in bacterial chemotaxis. Biochemistry. 1990 Jun 12;29(23):5436–5442. [PubMed]
  • Kuo SC, Koshland DE., Jr Multiple kinetic states for the flagellar motor switch. J Bacteriol. 1989 Nov;171(11):6279–6287. [PMC free article] [PubMed]

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