Global analysis of host targets of pathogen effector using yeast genetic array system

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Numerous bacterial pathogens of plants and animals utilize specialized secretion apparatus such as the type III secretion system to inject effector proteins directly into the host cell cytoplasm. Consequently, translocated effector proteins manipulate host cellular processes to promote infection and ultimately cause disease. In this study, we aimed to analyze the pathogen effectors from the plant pathogenic bacteria, Ralstonia solanacearum using yeast expression system and reveal the molecular mechanism of the strategy for R. solanacearum infection. Through the screening of 38 effectors from R. solanacearum GMI1000, we identified 5 effectors that caused sever growth inhibition to yeast cells. One of these effectors, REY4 protein, which has a domain conserved across phyla, was analyzed by the yeast genetic array system to identify the cellular target(s)in yeast. This screening identified that mutants having defect in the cell wall integrity MAP kinase cascade are hyper-sensitive to REY4-expression compared to wild-type cells. This result suggests that REY4 inhibits the pathway(s), which genetically interacts with the cell wall integrity MAP kinase cascade.