Carboxyl-terminal extensions beyond the conserved pentapeptide reduce rates of chemoreceptor adaptational modification.
Sensory adaptation in bacterial chemotaxis is mediated by covalent modification of chemoreceptors. Specific glutamyl residues are methylated and demethylated in reactions catalyzed by methyltransferase CheR and methylesterase CheB. In the well-characterized chemosensory systems of Escherichia coli and Salmonella spp., efficient modification by either enzyme is dependent on a conserved pentapeptide sequence, NWETF or NWESF, present at the extreme carboxyl terminus of high-abundance chemoreceptors. To what extent is position at the extreme carboxyl terminus important for pentapeptide-mediated enhancement of adaptational modification? Is this position equally important for enhancement of both enzyme activities? To address these questions, we created forms of high-abundance receptor Tsr or Tar carrying one, six, or eight additional amino acids extending beyond the pentapeptide at their carboxyl termini and assayed methylation, demethylation, deamidation, and ability to mediate chemotaxis. In vitro and in vivo, all three carboxyl-terminal extensions reduced pentapeptide-mediated enhancement of rates of adaptational modification. CheB-catalyzed reactions were more affected than CheR-catalyzed reactions. Effects were less severe for the complete sensory system in vivo than for the minimal system of receptor and modification enzymes in vitro. Notably, extended receptors mediated chemotaxis as efficiently as wild-type receptors, providing a striking example of robustness in chemotactic systems. This could reflect compensatory reductions of rates for both modification reactions, mitigation of effects of slower reactions by the intertwined circuitry of signaling and adaptation, or tolerance of a range of reactions rates for adaptational modification. No matter what the mechanism, the observations provide a challenging test for mathematical models of chemotaxis.
Lai
Wing-Cheung
WC
Department of Biochemistry, 117 Schweitzer Hall, University of Missouri-Columbia, Columbia, MI 65211, USA.
Hazelbauer
Gerald L
GL
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D002628:Chemoreceptor Cells / Q000737:chemistry / Q000378:metabolism*; D002633:Chemotaxis / Q000235:genetics / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology*; D008745:Methylation; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D012475:Salmonella / Q000235:genetics / Q000502:physiology*; D016415:Sequence Alignment; D015398:Signal Transduction / Q000235:genetics
D001426:Bacterial Proteins; D008780:Methyltransferases; C038706:chemotaxis methyltransferase; D002265:Carboxylic Ester Hydrolases; C041144:chemotactic protein methylesterase