Transmembrane signal transduction in bacterial chemotaxis involves ligand-dependent activation of phosphate group transfer.
Signal transduction in Escherichia coli involves the interaction of transmembrane receptor proteins such as the aspartate receptor, Tar, and the products of four chemotaxis genes, cheA, cheY, cheW, and cheZ. It was previously shown that the cheA gene product is an autophosphorylating protein kinase that transfers phosphate to CheY, whereas the cheZ gene product acts as a specific CheY phosphatase. Here we report that the system can be reconstituted in vitro and receptor function can be coupled to CheY phosphorylation. Coupling requires the presence of the CheW protein, the appropriate form of the receptor, and the CheA and CheY proteins. Under these conditions the accumulation of CheY-phosphate is enhanced approximately 300-fold. This rate enhancement is seen in reactions using wild-type and "tumble" mutant receptors but not "smooth" mutant receptors. The increased accumulation of phosphoprotein was inhibited by micromolar concentrations of aspartate, using wild-type, but not tumble, receptors. These results provide evidence that the signal transduction pathway in bacterial chemotaxis involves receptor-mediated alteration of the levels of phosphorylated proteins. They suggest that CheW acts as the coupling factor between receptor and phosphorylation. The results also support the suggestion that CheY-phosphate is the tumble signal.
Borkovich
K A
KA
Division of Biology, California Institute of Technology, Pasadena 91125.
Kaplan
N
N
Hess
J F
JF
Simon
M I
MI
D000255:Adenosine Triphosphate / Q000378:metabolism*; D015153:Blotting, Western; D002462:Cell Membrane / Q000502:physiology; D002630:Chemotactic Factors / Q000235:genetics* / Q000378:metabolism; D002633:Chemotaxis; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology*; D005798:Genes, Bacterial; D010766:Phosphorylation; D010957:Plasmids; D015398:Signal Transduction
D002630:Chemotactic Factors; D000255:Adenosine Triphosphate