title: Strong inhibition of fimbrial 3 subunit gene transcription by a novel downstream repressive element in Bordetella pertussis.
abstract: The Bvg-regulated promoters for the fimbrial subunit genes fim2 and fim3 of Bordetella pertussis behave differently from each other both in vivo and in vitro. In vivo Pfim2 is significantly stronger than Pfim3 , even though predictions based on the DNA sequences of BvgA-binding motifs and core promoter elements would indicate the opposite. In vitro Pfim3 demonstrated robust BvgA∼P-dependent transcriptional activation, while none was seen with Pfim2 . This apparent contradiction was investigated further. By swapping sequence elements we created a number of hybrid promoters and assayed their strength in vivo. We found that, while Pfim3 promoter elements upstream of the +1 transcriptional start site do indeed direct Bvg-activated transcription more efficiently than those of Pfim2 , the overall promoter strength of Pfim3  in vivo is reduced due to sequences downstream of +1 that inhibit transcription more than 250-fold. This element, the DRE (downstream repressive element), was mapped to the 15 bp immediately downstream of the Pfim3 +1. Placing the DRE in different promoter contexts indicated that its activity was not specific to fim promoters, or even to Bvg-regulated promoters. However it does appear to be specific to Bordetella species in that it did not function in Escherichia coli.
authors: Division of Bacterial, Parasitic, and Allergenic Products, Center for Biologics Evaluation and Research, FDA, Bethesda, MD, 20892, USA.
authors: Qing
authors: Q
authors: Chen
authors: Alice
authors: A
authors: Boulanger
authors: Deborah M
authors: DM
authors: Hinton
authors: Scott
authors: S
authors: Stibitz
mesh_terms: D000942:Antigens, Bacterial / Q000096:biosynthesis* / Q000235:genetics; D001886:Bordetella pertussis / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D036781:Fimbriae Proteins / Q000096:biosynthesis* / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D060847:Metabolic Engineering; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D050436:Regulatory Elements, Transcriptional; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D010566:Virulence Factors, Bordetella / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000942:Antigens, Bacterial; D010566:Virulence Factors, Bordetella; C101541:fim2 protein, Bordetella; C101542:fim3 protein, Bordetella; D036781:Fimbriae Proteins