Actin distribution is disrupted upon expression of Yersinia YopO/YpkA in yeast.
Pathogenic Yersinia spp. use a Type III secretion system to inject effector proteins directly into host cells. The effector proteins interfere with normal cellular signalling and disrupt cytoskeletal structures that are required for phagocytosis of the invader. Here we show that Saccharomyces cerevisiae responds to one of these effector proteins, YopO/YpkA, essentially as has been observed in mammalian model systems. Expression of YopO in yeast results in inhibition of growth on solid medium. YopO expression confers a cytotoxic effect upon yeast with only 6% of cells remaining viable after 3 h of expression. Moreover, there is no obvious cell cycle arrest associated with cytotoxicity. YopO expression disrupts the normal distribution of actin in yeast. Actin is no longer observed exclusively at sites of cell growth. Concurrent with disruption of actin, we find YopO localizes to the cell periphery, where it may disrupt actin localization. Inactivation of the YopO kinase has no apparent effect upon the deleterious effects of YopO. Collectively, our results show that the yeast model system that has been constructed meets all requirements to serve as an excellent genetic model to exploit examination of YopO and its intracellular targets.
Department of Biological Sciences, Western Michigan University, 1903 West Michigan Avenue, Kalamazoo, MI 49008, USA.
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Nejedlik
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Pierfelice
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Geiser
D000199:Actins / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D015870:Gene Expression; D005798:Genes, Bacterial; D008957:Models, Genetic; D010957:Plasmids / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D014170:Transformation, Genetic; D015007:Yersinia / Q000235:genetics*
D000199:Actins; D001426:Bacterial Proteins; D011994:Recombinant Proteins; C415206:YopO protein, Yersinia enterocolitica; C079893:ypkA protein, Yersinia; D017346:Protein Serine-Threonine Kinases