title: Only one of the five CheY homologs in Vibrio cholerae directly switches flagellar rotation.
abstract: Vibrio cholerae has three sets of chemotaxis (Che) proteins, including three histidine kinases (CheA) and four response regulators (CheY) that are encoded by three che gene clusters. We deleted the cheY genes individually or in combination and found that only the cheY3 deletion impaired chemotaxis, reinforcing the previous conclusion that che cluster II is involved in chemotaxis. However, this does not exclude the involvement of the other clusters in chemotaxis. In other bacteria, phospho-CheY binds directly to the flagellar motor to modulate its rotation, and CheY overexpression, even without CheA, causes extremely biased swimming behavior. We reasoned that a V. cholerae CheY homolog, if it directly controls flagellar rotation, should also induce extreme swimming behavior when overproduced. This was the case for CheY3 (che cluster II). However, no other CheY homolog, including the putative CheY (CheY0) protein encoded outside the che clusters, affected swimming, demonstrating that these CheY homologs cannot act directly on the flagellar motor. CheY4 very slightly enhanced the spreading of an Escherichia coli cheZ mutant in semisolid agar, raising the possibility that it can affect chemotaxis by removing a phosphoryl group from CheY3. We also found that V. cholerae CheY3 and E. coli CheY are only partially exchangeable. Mutagenic analyses suggested that this may come from coevolution of the interacting pair of proteins, CheY and the motor protein FliM. Taken together, it is likely that the principal roles of che clusters I and III as well as cheY0 are to control functions other than chemotaxis.
authors.affiliation: Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Hyakutake
authors.forename: Michio
authors.initials: M
authors.lastname: Homma
authors.forename: Melissa J
authors.initials: MJ
authors.lastname: Austin
authors.forename: Markus A
authors.initials: MA
authors.lastname: Boin
authors.forename: Claudia C
authors.initials: CC
authors.lastname: Häse
authors.forename: Ikuro
authors.initials: I
authors.lastname: Kawagishi
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D002633:Chemotaxis / Q000235:genetics* / Q000502:physiology; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology; D029968:Escherichia coli Proteins; D005407:Flagella / Q000235:genetics / Q000502:physiology*; D017353:Gene Deletion; D005816:Genetic Complementation Test; D000071677:Histidine Kinase; D008565:Membrane Proteins / Q000235:genetics; D000072236:Methyl-Accepting Chemotaxis Proteins; D008969:Molecular Sequence Data; D009068:Movement; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014734:Vibrio cholerae / Q000235:genetics / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; C470183:CheY-4 protein, Vibrio cholerae; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D000072236:Methyl-Accepting Chemotaxis Proteins; C000607606:cheY protein, E coli; D000071677:Histidine Kinase; C000607608:cheA protein, E coli