Use of phoA gene fusions to identify a pilus colonization factor coordinately regulated with cholera toxin.

RK Taylor, VL Miller, DB Furlong… - Proceedings of the …, 1987 - National Acad Sciences
RK Taylor, VL Miller, DB Furlong, JJ Mekalanos
Proceedings of the National Academy of Sciences, 1987National Acad Sciences
The transposon TnphoA was used to generate fusions between phoA, the gene for alkaline
phosphatase (PhoA), and genes encoding proteins that are secreted by Vibrio cholerae.
One of the PhoA+ mutants isolated showed a dramatic reduction in its ability to colonize the
intestines of suckling mice. This mutant no longer produced a 20.5-kDa protein (TcpA) that
we show is the major subunit of a V. cholerae pilus. Amino-terminal sequence analysis of
the TcpA pilus subunit showed that it shares amino acid homology with the pilins produced …
The transposon TnphoA was used to generate fusions between phoA, the gene for alkaline phosphatase (PhoA), and genes encoding proteins that are secreted by Vibrio cholerae. One of the PhoA+ mutants isolated showed a dramatic reduction in its ability to colonize the intestines of suckling mice. This mutant no longer produced a 20.5-kDa protein (TcpA) that we show is the major subunit of a V. cholerae pilus. Amino-terminal sequence analysis of the TcpA pilus subunit showed that it shares amino acid homology with the pilins produced by several other pathogenic bacteria. The TcpA pilus was coordinately expressed with cholera toxin under various culture conditions, and this effect appeared to be dependent on the transcriptional activator encoded by the toxR gene. We conclude that the toxR gene plays a central role in the transcriptional regulation of multiple virulence genes of V. cholerae.
National Acad Sciences