Mechanism of Signalling and Adaptation through the Rhodobacter sphaeroides Cytoplasmic Chemoreceptor Cluster

International Journal of Molecular Sciences

卷:20

JenniferA. de Beyer¹  Elaine Byles¹  JudithP. Armitage¹  Andrea Szöllössi¹  Roman Fischer² 

[1] Department of Biochemistry, University of Oxford, Oxford OX1 3QU, UK;

[2] Nuffield Department of Medicine, University of Oxford, Oxford OX3 9DU, UK;

关键词: chemotaxis;    rhodobacter;    adaptation;    chemoreceptors;    cytoplasmic signalling;    mass spectrometry;    signal transduction;    signal integration;    methyl-accepting chemotaxis proteins;   

DOI  :  10.3390/ijms20205095

来源: DOAJ

【 摘 要 】

Rhodobacter sphaeroides has two chemotaxis clusters, an Escherichia coli-like cluster with membrane-spanning chemoreceptors and a less-understood cytoplasmic cluster. The cytoplasmic CheA is split into CheA₄, a kinase, and CheA₃, a His-domain phosphorylated by CheA₄ and a phosphatase domain, which together phosphorylate and dephosphorylate motor-stopping CheY₆. In bacterial two-hybrid analysis, one major cytoplasmic chemoreceptor, TlpT, interacted with CheA₄, while the other, TlpC, interacted with CheA₃. Both clusters have associated adaptation proteins. Deleting their methyltransferases and methylesterases singly and together removed chemotaxis, but with opposite effects. The cytoplasmic cluster signal overrode the membrane cluster signal. Methylation and demethylation of specific chemoreceptor glutamates controls adaptation. Tandem mass spectroscopy and bioinformatics identified four putative sites on TlpT, three glutamates and a glutamine. Mutating each glutamate to alanine resulted in smooth swimming and loss of chemotaxis, unlike similar mutations in E. coli chemoreceptors. Cells with two mutated glutamates were more stoppy than wild-type and responded and adapted to attractant addition, not removal. Mutating all four sites amplified the effect. Cells were non-motile, began smooth swimming on attractant addition, and rapidly adapted back to non-motile before attractant removal. We propose that TlpT responds and adapts to the cell’s metabolic state, generating the steady-state concentration of motor-stopping CheY₆~P. Membrane-cluster signalling produces a pulse of CheY₃/CheY₄~P that displaces CheY₆~P and allows flagellar rotation and smooth swimming before both clusters adapt.

【 授权许可】

Unknown