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Journal of Bacteriology, February 2000, p. 967-973, Vol. 182, No. 4
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Polar Clustering of the Chemoreceptor Complex in Escherichia coli Occurs in the Absence of Complete CheA Function

J. M. Skidmore,1 D. D. Ellefson,2,dagger B. P. McNamara,2,Dagger M. M. P. Couto,1 A. J. Wolfe,2 and J. R. Maddock1,*

Department of Biology, University of Michigan, Ann Arbor, Michigan 48109-1048,1 and Department of Microbiology and Immunology, Loyola University Chicago, Maywood, Illinois 601532

Received 2 September 1999/Accepted 19 November 1999

Bacterial chemotaxis requires a phosphorelay system initiated by the interaction of a ligand with its chemoreceptor and culminating in a change in the directional bias of flagellar rotation. Chemoreceptor-CheA-CheW ternary complexes mediate transduction of the chemotactic signal. In vivo, these complexes cluster predominantly in large groups at the cell poles. The function of chemoreceptor clustering is currently unknown. To gain insight into the relationship between signaling and chemoreceptor clustering, we examined these properties in several Escherichia coli mutant strains that produce CheA variants altered in their ability to mediate chemotaxis, autophosphorylate, or bind ATP. We show here that polar clustering of chemoreceptor complexes does not require functional CheA protein, although maximal clustering occurred only in chemotactically competent cells. Surprisingly, in cells containing a minimum of 13 gold particles at the cell pole, a significant level of clustering was observed in the absence of CheA, demonstrating that CheA is not absolutely essential for chemoreceptor clustering. Nonchemotactic cells expressing only CheAS, a C-terminal CheA deletion, or CheA bearing a mutation in the ATP-binding site mediated slightly less than maximal chemoreceptor clustering. Cells expressing only full-length CheA (CheAL) from either a chromosomal or a plasmid-encoded allele displayed a methyl-accepting chemotaxis protein localization pattern indistinguishable from that of strains carrying both CheAL and CheAS, demonstrating that CheAL alone can mediate polar clustering.

* Corresponding author. Mailing address: Department of Biology, University of Michigan, 830 North University, Ann Arbor, MI 48109-1048. Phone: (734) 936-8068. Fax: (734) 647-0884. E-mail: maddock{at}umich.edu.

dagger Present address: Department of Molecular Microbiology and Immunology, Oregon Health Sciences University, Portland, OR 97201.

Dagger Present address: Infectious Diseases, University of Maryland, Baltimore, MD 21201.

Journal of Bacteriology, February 2000, p. 967-973, Vol. 182, No. 4
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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