987P fimbrial gene identification and protein characterization by T7 RNA polymerase-induced transcription and TnphoA mutagenesis.
The 987P fimbrial gene cluster has been previously cloned as a 12 kb fragment from prototype strain 987. Gene products encoded by the whole clone were analysed by utilizing an in vivo system based on the induction of transcription by T7 RNA polymerase. The sensitivity of this technique permitted us to identify new proteins involved in 987P fimbriation. In total, eight proteins were detected, their genes (fasA to fasH) were mapped and their orientation of transcription determined. Several of the gene products demonstrated typical properties of exported proteins. Precursor and processed forms could be correlated after inhibiting protein transport with ethanol. The detection of enzymatically active fusion proteins after TnphoA (Tn5IS50L::phoA) mutagenesis supported and complemented these results. One protein encoded by the 12kb fragment was found not to be related to fimbriation but rather the product of the STla gene, identified as a component of a Tn1681-like transposon.
Schifferli
D M
DM
Department of Microbiology and Immunology, University of Tennessee, Memphis 38104.
Beachey
E H
EH
Taylor
R K
RK
D018830:Adhesins, Escherichia coli; D000942:Antigens, Bacterial / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics; D001427:Bacterial Toxins / Q000235:genetics; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004768:Enterotoxins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial; D005798:Genes, Bacterial; D008040:Genetic Linkage; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014764:Viral Proteins
D018830:Adhesins, Escherichia coli; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004251:DNA Transposable Elements; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D014764:Viral Proteins; C035391:antigen 987P, E coli; C034151:heat stable toxin (E coli); D036781:Fimbriae Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases