J. Bacteriol., 07 1996, 4208-4215, Vol 178, No. 14
Copyright © 1996, American Society for Microbiology

Tyrosine 106 of CheY plays an important role in chemotaxis signal transduction in Escherichia coli

X Zhu, CD Amsler, K Volz and P Matsumura
Department of Microbiology and Immunology, University of Illinois at Chicago, 60612-7344, USA.

CheY is the response regulator in the signal transduction pathway of bacterial chemotaxis. Position 106 of CheY is occupied by a conserved aromatic residue (tyrosine or phenylalanine) in the response regulator superfamily. A number of substitutions at position 106 have been made and characterized by both behavioral and biochemical studies. On the basis of the behavioral studies, the phenotypes of the mutants at position 106 can be divided into three categories: (i) hyperactivity, with a tyrosine-to-tryptophan mutation (Y106W) causing increased tumble signaling but impairing chemotaxis; (ii) low-level activity, with a tyrosine-to-phenylalanine change (Y106F) resulting in decreased tumble signaling and chemotaxis; and (iii) no activity, with substitutions such as Y106L, Y106I, Y106V, Y106G, and Y106C resulting in no chemotaxis and a smooth-swimming phenotype. All three types of mutants can be phosphorylated by CheA-phosphate in vitro to a level similar to that of wild-type CheY. Autodephosphorylation rates are similar for all categories of mutants. All mutant proteins displayed less than twofold increased rates compared with wild-type CheY. Binding of the mutant proteins to FliM was similar to that of the wild-type CheY in the CheY- FliM binding assays. The combined results from in vivo behavioral and in vitro biochemical studies suggest that the diverse phenotypes of the Y106 mutants are not due to a variation in phosphorylation or dephosphorylation ability nor in affinity for the switch. With reference to the structures of wild-type CheY and the T871 CheY mutant, our results suggest that rearrangements of the orientation of the tyrosine side chain at position 106 are involved in the signal transduction of CheY. These data also suggest that the binding of phosphoryl-CheY to the flagellar motor is a necessary, but not sufficient, event for signal transduction.

This article has been cited by other articles:

• Caballero-Manrique, E., Bray, J. K., Deutschman, W. A., Dahlquist, F. W., Guenza, M. G. (2007). A Theory of Protein Dynamics to Predict NMR Relaxation. Biophys. J 93: 4128-4140 [Abstract] [Full Text]  
• Knaggs, M. H., Salsbury, F. R. Jr., Edgell, M. H., Fetrow, J. S. (2007). Insights into Correlated Motions and Long-Range Interactions in CheY Derived from Molecular Dynamics Simulations. Biophys. J 92: 2062-2079 [Abstract] [Full Text]  
• Varughese, K. I. (2005). Conformational Changes of Spo0F along the Phosphotransfer Pathway. J. Bacteriol. 187: 8221-8227 [Abstract] [Full Text]  
• Ferre, A., de la Mora, J., Ballado, T., Camarena, L., Dreyfus, G. (2004). Biochemical Study of Multiple CheY Response Regulators of the Chemotactic Pathway of Rhodobacter sphaeroides. J. Bacteriol. 186: 5172-5177 [Abstract] [Full Text]  
• Szurmant, H., Ordal, G. W. (2004). Diversity in Chemotaxis Mechanisms among the Bacteria and Archaea. Microbiol. Mol. Biol. Rev. 68: 301-319 [Abstract] [Full Text]  
• Butler, S. M., Camilli, A. (2004). Both chemotaxis and net motility greatly influence the infectivity of Vibrio cholerae. Proc. Natl. Acad. Sci. USA 101: 5018-5023 [Abstract] [Full Text]  
• Roche, P., Mouawad, L., Perahia, D., Samama, J.-P., Kahn, D. (2002). Molecular dynamics of the FixJ receiver domain: movement of the {beta}4-{alpha}4 loop correlates with the in and out flip of Phe101. Protein Sci. 11: 2622-2630 [Abstract] [Full Text]  
• Re, S. D., Tolstykh, T., Wolanin, P. M., Stock, J. B. (2002). Genetic analysis of response regulator activation in bacterial chemotaxis suggests an intermolecular mechanism. Protein Sci. 11: 2644-2654 [Abstract] [Full Text]  
• Guillet, V., Ohta, N., Cabantous, S., Newton, A., Samama, J.-P. (2002). Crystallographic and Biochemical Studies of DivK Reveal Novel Features of an Essential Response Regulator in Caulobacter crescentus. J. Biol. Chem. 277: 42003-42010 [Abstract] [Full Text]  
• Im, Y. J., Rho, S.-H., Park, C.-M., Yang, S.-S., Kang, J.-G., Lee, J. Y., Song, P.-S., Eom, S. H. (2002). Crystal structure of a cyanobacterial phytochrome response regulator. Protein Sci. 11: 614-624 [Abstract] [Full Text]  
• Qin, L., Yoshida, T., Inouye, M. (2001). The critical role of DNA in the equilibrium between OmpR and phosphorylated OmpR mediated by EnvZ in Escherichiacoli. Proc. Natl. Acad. Sci. USA 10.1073/pnas.031383098v1 [Abstract] [Full Text]  
• Bren, A., Eisenbach, M. (2000). How Signals Are Heard during Bacterial Chemotaxis: Protein-Protein Interactions in Sensory Signal Propagation. J. Bacteriol. 182: 6865-6873 [Full Text]  
• Scharf, B. E., Fahrner, K. A., Berg, H. C. (1998). CheZ Has No Effect on Flagellar Motors Activated by CheY13DK106YW. J. Bacteriol. 180: 5123-5128 [Abstract] [Full Text]  
• Appleby, J. L., Bourret, R. B. (1998). Proposed Signal Transduction Role for Conserved CheY Residue Thr87, a Member of the Response Regulator Active-Site Quintet. J. Bacteriol. 180: 3563-3569 [Abstract] [Full Text]  
• McEvoy, M. M., Hausrath, A. C., Randolph, G. B., Remington, S. J., Dahlquist, F. W. (1998). Two binding modes reveal flexibility in kinase/response regulator interactions in the bacterial chemotaxis pathway. Proc. Natl. Acad. Sci. USA 95: 7333-7338 [Abstract] [Full Text]  
• Ramakrishnan, R., Schuster, M., Bourret, R. B. (1998). Acetylation at Lys-92 enhances signaling by the chemotaxis response regulator protein CheY. Proc. Natl. Acad. Sci. USA 95: 4918-4923 [Abstract] [Full Text]  
• Elliot, M., Damji, F., Passantino, R., Chater, K., Leskiw, B. (1998). The bldD Gene of Streptomyces coelicolor A3(2): a Regulatory Gene Involved in Morphogenesis and Antibiotic Production. J. Bacteriol. 180: 1549-1555 [Abstract] [Full Text]  
• Scharf, B. E., Fahrner, K. A., Turner, L., Berg, H. C. (1998). Control of direction of flagellar rotation in bacterial chemotaxis. Proc. Natl. Acad. Sci. USA 95: 201-206 [Abstract] [Full Text]  
• Zhu, X., Volz, K., Matsumura, P. (1997). The CheZ-binding Surface of CheY Overlaps the CheA- and FliM-binding Surfaces. J. Biol. Chem. 272: 23758-23764 [Abstract] [Full Text]  
• Jiang, M., Bourret, R. B., Simon, M. I., Volz, K. (1997). Uncoupled Phosphorylation and Activation in Bacterial Chemotaxis. THE 2.3 A STRUCTURE OF AN ASPARTATE TO LYSINE MUTANT AT POSITION 13 OF CheY. J. Biol. Chem. 272: 11850-11855 [Abstract] [Full Text]  
• Zhu, X., Rebello, J., Matsumura, P., Volz, K. (1997). Crystal Structures of CheY Mutants Y106W and T87I/Y106W. CheY ACTIVATION CORRELATES WITH MOVEMENT OF RESIDUE 106. J. Biol. Chem. 272: 5000-5006 [Abstract] [Full Text]  
• Simonovic, M., Volz, K. (2001). A Distinct Meta-active Conformation in the 1.1-A Resolution Structure of Wild-type ApoCheY. J. Biol. Chem. 276: 28637-28640 [Abstract] [Full Text]  
• Cervin, M. A., Spiegelman, G. B. (2000). A Role for Asp75 in Domain Interactions in the Bacillus subtilis Response Regulator Spo0A. J. Biol. Chem. 275: 22025-22030 [Abstract] [Full Text]  
• Lee, S.-Y., Cho, Ho. S., Pelton, J. G., Yan, D., Berry, E. A., Wemmer, D. E. (2001). Crystal Structure of Activated CheY. COMPARISON WITH OTHER ACTIVATED RECEIVER DOMAINS. J. Biol. Chem. 276: 16425-16431 [Abstract] [Full Text]  
• Schuster, M., Zhao, R., Bourret, R. B., Collins, E. J. (2000). Correlated Switch Binding and Signaling in Bacterial Chemotaxis. J. Biol. Chem. 275: 19752-19758 [Abstract] [Full Text]  
• Qin, L., Yoshida, T., Inouye, M. (2001). The critical role of DNA in the equilibrium between OmpR and phosphorylated OmpR mediated by EnvZ in Escherichiacoli. Proc. Natl. Acad. Sci. USA 98: 908-913 [Abstract] [Full Text]  
• Schuster, M., Silversmith, R. E., Bourret, R. B. (2001). Conformational coupling in the chemotaxis response regulator CheY. Proc. Natl. Acad. Sci. USA 98: 6003-6008 [Abstract] [Full Text]