Genetic evidence that the yopA gene-encoded Yersinia outer membrane protein Yop1 mediates inhibition of the anti-invasive effect of interferon.
HEp-2 cell monolayers were challenged with genetic variants of Yersinia pseudotuberculosis YPIII(pIB1) and Yersinia enterocolitica W22708(pYL4). Both strains were represented by (i) variants harboring the 70-kilobase virulence plasmid, (ii) their isogenic plasmid-cured derivatives, and (iii) two transposon mutants constructed by insertional inactivation of the plasmid genes encoding outer membrane protein Yop1 and Ca2+ dependency in strains YPIII(pIB1) and W22708(pYL4). When the HEp-2 cells were pretreated with recombinant alpha interferon subtype A, all invasive variants of Y. enterocolitica and Y. pseudotuberculosis, except those variants which expressed Yop1, showed a significantly reduced ability to localize intracellularly. The anti-invasive effect of interferon was abolished when the gene was expressed or when a sterile filtered sonic extract of a Yop1-producing strain was added to the cell cultures. To obtain further evidence of a potential role of Yop1, a DNA fragment encoding Yop1 cloned into the vector pBR322 was used. After introduction of the resultant recombinant plasmid pYMS2 into the plasmid-cured variant YPIII and the Yop1-negative transposon mutant YPIII(pIB102) of Y. pseudotuberculosis, both transformants regained the ability to produce Yop1 and showed complete inhibition of the interferon effect. Moreover, the sterile sonic extract of an Escherichia coli strain, which carried pYMS2, neutralized the anti-invasive effect of interferon. The results provide direct genetic evidence that Yop1 mediates inhibition of the anti-invasive effect of interferon in HEp-2 cell cultures. The results also demonstrated that Yop1 itself reduces the ability of Yersinia spp. to localize intracellularly in HEp-2 cells.
Bukholm
G
G
Kaptein W. Wilhelmsen og Frues Bakteriologiske Institutt, University of Oslo, Norway.
Kapperud
G
G
Skurnik
M
M
D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000276:immunology*; D002460:Cell Line; D003001:Cloning, Molecular; D006801:Humans; D007370:Interferon Type I / Q000502:physiology*; D010957:Plasmids; D012486:Salmonella typhimurium / Q000276:immunology; D014170:Transformation, Genetic; D015008:Yersinia enterocolitica / Q000472:pathogenicity*; D015011:Yersinia pseudotuberculosis / Q000472:pathogenicity*
D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D007370:Interferon Type I; C064832:YadA protein, Yersinia