Expression of Bordetella pertussis fimbrial (fim) genes in Bordetella bronchiseptica: fimX is expressed at a low level and vir-regulated.
To identify the elements involved in Bordetella pertussis fimbriae regulation and to determine whether fimX is an expressed gene, the promoter regions of fimX, fim2 and fim3 from strain BPSA1 were isolated and linked to the promoterless CAT gene in pLAFR2. By following CAT activity in Bordetella bronchiseptica vir+ and vir- strains we established that the fimX promoter, like those for fim2 and fim3, is active, although at a low level, and vir-regulated. This suggests that the fimX protein might be produced in minute quantities which are not detectable by conventional methods. Comparison of the three fim promoter sequences and transcriptional activities identifies two conserved elements necessary for transcription in the -60 to -20 region: the 'fim box' and the 'C-stretch'. Mutations in these two sequences drastically reduce transcription and alter the interaction with vir components, suggesting a role for the two elements in the regulation of fim genes. Finally, we suggest that the apparent constitutive nature of fim3 in BPSA1 is due to a modification in the length of the 'C-stretch'.
Eniricerche S.p.A., Department of Genetic Engineering and Microbiology, Milan, Italy.
B
B
Riboli
P
P
Pedroni
A
A
Cuzzoni
G
G
Grandi
F
F
de Ferra
D001483:Base Sequence; D001884:Bordetella / Q000235:genetics*; D016950:Bordetella bronchiseptica / Q000235:genetics; D001886:Bordetella pertussis / Q000235:genetics; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D004277:DNA, Single-Stranded / Q000235:genetics; D010861:Fimbriae, Bacterial / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014158:Transcription, Genetic
D004277:DNA, Single-Stranded; D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase