Transcription of the yop regulon from Y. enterocolitica requires trans acting pYV and chromosomal genes

Virulent Y. enterocolitica strains restrict their growth at 37 degrees C, in rich medium deprived of calcium. This property, called calcium dependency, correlates with pathogenicity. It is conditioned by a 70 kb plasmid called pYV. The analysis of calcium independent (Cl) insertion mutants defined a 20 kb region called the calcium region. This region contains at least three transcription units called virA, B and C. In growth restriction conditions, Y. enterocolitica releases and inserts in its outer-membrane several pYV encoded proteins (POMPs). By transposition mutagenesis using a mini-Mu dlac element, we localized plasmid genes encoding POMPs of 84, 51, 30 and 25 kDa. These genes appear to be scattered on pYV, outside the calcium region and to constitute a regulon. Transcription of these yop genes increased by factors of 3.5 (yop84) to 200 (yop51) when temperature was shifted from 25 to 37 degrees C. On the contrary, calcium had only a moderate effect. Transcription was also dependent on the culture medium and on a trans-acting factor encoded by the calcium region. One Cl double mutant severely affected in transcription of yop51 defined a new locus called virF in the calcium region of pYV. Transcription of the yop genes was very poor in E. coli K12, even in the presence of a fully functional calcium region.