Mutational analysis of the chemoreceptor-coupling domain of the Escherichia coli chemotaxis signaling kinase CheA.

Journal of Bacteriology
Jinshi Zhao, John S Parkinson

Abstract

During chemotactic signaling by Escherichia coli, autophosphorylation of the histidine kinase CheA is coupled to chemoreceptor control by the CheW protein, which interacts with the C-terminal P5 domain of CheA. To identify P5 determinants important for CheW binding and receptor coupling control, we isolated and characterized a series of P5 missense mutants. The mutants fell into four phenotypic groups on the basis of in vivo behavioral and protein stability tests and in vitro assays with purified mutant proteins. Group 1 mutants exhibited autophosphorylation and receptor-coupling defects, and their CheA proteins were subject to relatively rapid degradation in vivo. Group 1 mutations were located at hydrophobic residues in P5 subdomain 2 and most likely caused folding defects. Group 2 mutants made stable CheA proteins with normal autophosphorylation ability but with defects in CheW binding and in receptor-mediated activation of CheA autophosphorylation. Their mutations affected residues in P5 subdomain 1 near the interface with the CheA dimerization (P3) and ATP-binding (P4) domains. Mutant proteins of group 3 were normal in all tests yet could not support chemotaxis, suggesting that P5 has one or more important but still unknow...Continue Reading

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Citations

Feb 23, 2012·Proceedings of the National Academy of Sciences of the United States of America·Ariane BriegelBrian R Crane
May 5, 2012·Proceedings of the National Academy of Sciences of the United States of America·Jun LiuWilliam Margolin
May 29, 2010·The Journal of Biological Chemistry·Felipe TrajtenbergAlejandro Buschiazzo
Jul 8, 2009·Journal of Bacteriology·Christopher O'ConnorAndres Campos
Oct 4, 2011·Journal of Bacteriology·Hai The Pham, John S Parkinson
Jan 1, 2014·Proceedings of the National Academy of Sciences of the United States of America·Simon RinggaardMatthew K Waldor
Jul 22, 2006·Molecular Microbiology·David KentnerVictor Sourjik
Jun 9, 2010·Molecular Microbiology·Kathryn A ScottJudith P Armitage
Jun 5, 2012·Journal of Molecular Biology·Xiqing WangFrederick W Dahlquist
Apr 4, 2015·Trends in Microbiology·John S ParkinsonJoseph J Falke
Jul 25, 2012·Biophysical Journal·Jaya BhatnagarBrian R Crane
Nov 29, 2017·Molecular Plant-microbe Interactions : MPMI·Zhiwei HuangMinliang Guo
Feb 28, 2018·Journal of Bacteriology·Xueye DingXiqing Wang
Mar 14, 2018·Journal of Bacteriology·Simon RinggaardAriane Briegel
Apr 20, 2006·Nature Structural & Molecular Biology·Sang-Youn ParkBrian R Crane

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