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Journal of Bacteriology, January 1999, p. 627-631, Vol. 181, No. 2
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Dissecting the VanRS Signal Transduction Pathway with Specific Inhibitors

Andrew T. Ulijasz and Bernard Weisblum*

Department of Pharmacology, University of Wisconsin Medical School, Madison, Wisconsin 53706

Received 2 September 1998/Accepted 30 October 1998

The VanRS two-component signal transduction pathway from Enterococcus faecium was reconstituted in vitro from partially purified components and shown to be inhibited by the halophenyl isothiazolone LY-266,400, inhibitor A, a compound shown previously to reduce expression of the AlgR1-AlgR2 two-component signal transduction pathway in Pseudomonas aeruginosa (S. Roychoudhury, N. A. Zielinski, A. J. Ninfa, N. E. Allen, L. N. Jungheim, T. I. Nicas, and A. M. Chakrabarty, Proc. Natl. Acad. Sci. USA 90:965-969, 1993). Inhibitor A attenuates phosphoryl transfer from VanS~P to VanR by its action on the ability of VanR to accept. We observed an apparent stimulatory effect of inhibitor A on VanS autophosphorylation which is attributable to the accumulation of VanS~P as an intermediate unable to transfer Pi to the inhibited VanR. Thus, inhibitor A acts on the second of two sequential steps which lead to transcriptional activation of the VanHAXYZ gene cluster and the resultant expression of vancomycin resistance.

* Corresponding author. Mailing address: Department of Pharmacology, University of Wisconsin Medical School, 1300 University Ave., Madison, WI 53706. Phone: (608) 262-0972. Fax: (608) 262-1257. E-mail: weisblum{at}macc.wisc.edu.

Journal of Bacteriology, January 1999, p. 627-631, Vol. 181, No. 2
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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