Characterization of a novel fucose-regulated promoter (PfcsK) suitable for gene essentiality and antibacterial mode-of-action studies in Streptococcus pneumoniae.
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 P(fcsK), fucose-dependent growth of the S. pneumoniae (P(fcsK)::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.
Chan
Pan F
PF
Microbial, Musculoskeletal and Proliferative Diseases Center of Excellence for Drug Discovery, GlaxoSmithKline Pharmaceuticals, Collegeville, Pennsylvania 19426, USA. Pan_2_Chan@gsk.com
O'Dwyer
Karen M
KM
Palmer
Leslie M
LM
Ambrad
Jennifer D
JD
Ingraham
Karen A
KA
So
Chi
C
Lonetto
Michael A
MA
Biswas
Sanjoy
S
Rosenberg
Martin
M
Holmes
David J
DJ
Zalacain
Magdalena
M
D000581:Amidohydrolases; D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000235:genetics; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005643:Fucose / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D020043:Genes, Essential; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D008969:Molecular Sequence Data; D009876:Operon; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013296:Streptococcus pneumoniae / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D006877:Hydroxamic Acids; D005643:Fucose; D000626:Aminopeptidases; D000581:Amidohydrolases; C084507:peptide deformylase; C010531:actinonin