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Journal of Bacteriology, March 2003, p. 2051-2058, Vol. 185, No. 6
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.6.2051-2058.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Characterization of a Novel Fucose-Regulated Promoter (PfcsK) Suitable for Gene Essentiality and Antibacterial Mode-of-Action Studies in Streptococcus pneumoniae

Pan F. Chan,* Karen M. O'Dwyer, Leslie M. Palmer, Jennifer D. Ambrad, Karen A. Ingraham, Chi So, Michael A. Lonetto, Sanjoy Biswas, Martin Rosenberg,{dagger} David J. Holmes, and Magdalena Zalacain

Microbial, Musculoskeletal and Proliferative Diseases Center of Excellence for Drug Discovery, GlaxoSmithKline Pharmaceuticals, Collegeville, Pennsylvania 19426

Received 11 September 2002/ Accepted 18 December 2002

The promoter of the Streptococcus pneumoniae putative fuculose kinase gene (fcsK), the first gene of a novel fucose utilization operon, is induced by fucose and repressed by glucose or sucrose. When the streptococcal polypeptide deformylase (PDF) gene (def1, encoding PDF) was placed under the control of PfcsK, fucose-dependent growth of the S. pneumoniae (PfcsK::def1) strain was observed, confirming the essential nature of PDF in this organism. The mode of antibacterial action of actinonin, a known PDF inhibitor, was also confirmed with this strain. The endogenous fuculose kinase promoter is a tightly regulated, titratable promoter which will be useful for target validation and for confirmation of the mode of action of novel antibacterial drugs in S. pneumoniae.

* Corresponding author. Mailing address: GlaxoSmithKline Pharmaceuticals, Microbial, Musculoskeletal and Proliferative Diseases CEDD, 1250 South Collegeville Rd., P.O. Box 5089, UP1345, Collegeville, PA 19426-0989. Phone: (610) 917-5255. Fax: (610) 917-7901. E-mail: Pan_2_Chan{at}gsk.com.

{dagger} Present address: Promega Corporation, Madison, WI 53711.

Journal of Bacteriology, March 2003, p. 2051-2058, Vol. 185, No. 6
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.6.2051-2058.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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