title: Roles of the highly conserved aspartate and lysine residues in the response regulator of bacterial chemotaxis.
abstract: The chemotactic responses of bacteria such as Escherichia coli and Salmonella typhimurium are mediated by phosphorylation of the CheY protein. Phospho-CheY interacts with the flagellar motor switch to cause tumbly behavior. CheY belongs to a large family of phosphorylated response regulators that function in bacteria to control motility and regulate gene expression. Residues corresponding to Asp57, Asp13, and Lys109 in CheY are highly conserved among all of these proteins. The site of phosphorylation in CheY is Asp57, and in the three-dimensional structure of CheY the Asp57 carboxylate side chain is in close proximity to the beta-carboxylate of Asp13 and the epsilon-amin of Lys109. To further examine the roles of these residues in response regulator function, each has been mutated to a conservative substitution. Asn for Asp and Arg for Lys. All mutations abolished CheY function in vivo. Whereas the Asp to Asn mutations dramatically reduced levels of CheY phosphorylation, the Lys to Arg mutation had the opposite effect. The high level of phosphorylation in the Lys109 mutant results from a decreased autophosphatase activity as well as a lack of phosphatase stimulation by the phosphatase activating protein, CheZ. Despite its high level of phosphorylation, the Lys109 mutant protein cannot produce tumbly behavior. Thus, Lys109 is required for an event subsequent to phosphorylation. We propose that an interaction between the epsilon-amino of Lys109 and the phosphoryl group at Asp57 is essential for the conformational switch that leads to activation of CheY.
authors.affiliation: Department of Molecular Biology, Princeton University, New Jersey 08544-1014.
authors.forename: G S
authors.initials: GS
authors.lastname: Lukat
authors.forename: B H
authors.initials: BH
authors.lastname: Lee
authors.forename: J M
authors.initials: JM
authors.lastname: Mottonen
authors.forename: A M
authors.initials: AM
authors.lastname: Stock
authors.forename: J B
authors.initials: JB
authors.lastname: Stock
mesh_terms: D001224:Aspartic Acid / Q000502:physiology*; D001426:Bacterial Proteins; D002630:Chemotactic Factors / Q000378:metabolism*; D002633:Chemotaxis / Q000502:physiology*; D029968:Escherichia coli Proteins; D005407:Flagella / Q000378:metabolism; D008239:Lysine / Q000502:physiology*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D000072236:Methyl-Accepting Chemotaxis Proteins; D009154:Mutation; D010766:Phosphorylation; D012486:Salmonella typhimurium / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D002630:Chemotactic Factors; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D000072236:Methyl-Accepting Chemotaxis Proteins; C000607606:cheY protein, E coli; D001224:Aspartic Acid; D008239:Lysine