Analysis of the yopA gene encoding the Yop1 virulence determinants of Yersinia spp.

Abstract

The Yop proteins of Yersinia are important virulence determinants. The Yop1 protein sequences of Yersinia pestis, Yersinia pseudotuberculosis, and two Yersinia enterocolitica serotypes, 0:3 and 0:8, deduced from the nucleotide sequences of the corresponding yopA genes, were compared. Most differences were found in the hydrophilic domains of the proteins, whereas the hydrophobic domains were conserved. The amino acid sequences revealed a signal sequence 25 amino acids long. No cysteine residues were present, even though Yop1 forms a polymeric structure. The transcription startpoint of yopA was determined by primer extension. The coding region and transcription startpoint were separated by a leader sequence 270 nucleotides long. The yopA promoter sequence of Y.pestis is identical to the corresponding sequence of Y. pseudotuberculosis and transcription studies revealed that this promoter is active in Y.pestis. Thus, the inability of Y.pestis to express the Yop1 protein is due to a single base pair deletion in the coding region of the yopA gene of Y.pestis.

LcrF is the temperature-regulated activator of the yadA gene of Yersinia enterocolitica and Yersinia pseudotuberculosis

“…Because of a 1-bp deletion early in the yadA gene, it is not expressed in Y. pestis (31). In addition, YadA expression is affected by the growth medium such that highest expression is achieved in a poor medium (16).…”

“…In addition, YadA expression is affected by the growth medium such that highest expression is achieved in a poor medium (16). The yadA gene has been sequenced, and the transcription start point has been determined (31). The open reading frame and transcription start are separated by a 270-nucleotide-long leader sequence (31), which may play a role in the temperature regulation of YadA expression.…”

“…The yadA gene has been sequenced, and the transcription start point has been determined (31). The open reading frame and transcription start are separated by a 270-nucleotide-long leader sequence (31), which may play a role in the temperature regulation of YadA expression. Moreover, the promoter region of theyadA gene is very A+T rich, a phenomenon typical of DNA regions where regulatory proteins bind after temperature-induced topological changes have taken place.…”

“…To determine whether the IcrLMC locus of pYV (14) contains the regulatory genes of yadA expression, we constructed a strain carrying the yadA4 gene and the lcrLMC locus in trans. Plasmids carrying the lcrLMC locus were isolated from the ClaI libraries of pYV8081, pIB1, and pYVO3 (4,31). From the known location of lcrLMC and the restriction map of pYVO3 (23) (Fig.…”

Application of the polymerase chain reaction and immunofluorescence techniques to the detection of bacteria in Yersinia‐triggered reactive arthritis

“…DNA was spotted onto the filters with a vacuumdriven 96-well apparatus, and the filters were baked for 2 hours at 80°C. Prehybridization was performed in 1 x Denhardt's solution (0.02% polyvinylpyrrolidone, 0.02% Ficoll, and 0.02% bovine serum albumin), 1% sodium dodecyl sulfate (SDS), and 6 x SSC for 2 hours at 65"C, and hybridization was performed in l x Denhardt's solution, 1% SDS, 6 x SSC, and 100 pg/ml of single-stranded DNA (ssDNA) at 65°C for [16][17][18] hours.…”