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Biochemical Study of Multiple CheY Response Regulators of the Chemotactic Pathway of Rhodobacter sphaeroides

Axelle Ferré,^1, Javier de la Mora,^1, Teresa Ballado,^1, Laura Camarena,² and Georges Dreyfus^1*

Departamento de Genética Molecular, Instituto de Fisiología Celular,¹ Departamento de Biología Molecular, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, 04510 Mexico DF, Mexico²

Received 3 March 2004/ Accepted 22 April 2004

The six copies of the response regulator CheY from Rhodobacter sphaeroides bind to the switch protein FliM. Phosphorylation by acetyl phosphate (AcP) was detected by tryptophan fluorescence quenching in three of the four CheYs that contain this residue. Autophosphorylation with Ac³²P was observed in five CheY proteins. We also show that all of the cheY genes are expressed simultaneously; therefore, in vivo all of the CheY proteins could bind to FliM to control the chemotactic response. Consequently, we hypothesize that in this complex chemotactic system, the binding of some CheY proteins to FliM, does not necessarily imply switching of the flagellar motor.

This study is dedicated to the memory of our dear friend Robert M. Macnab.

A.F., J.D.L.M., and T.B. contributed equally to this study.

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