title: Expression of Pseudomonas syringae type III effectors in yeast under stress conditions reveals that HopX1 attenuates activation of the high osmolarity glycerol MAP kinase pathway.
abstract: The Gram-negative bacterium Pseudomonas syringae pv. tomato (Pst) is the causal agent of speck disease in tomato. Pst pathogenicity depends on a type III secretion system that delivers effector proteins into host cells, where they promote disease by manipulating processes to the advantage of the pathogen. Previous studies identified seven Pst effectors that inhibit growth when expressed in yeast under normal growth conditions, suggesting that they interfere with cellular processes conserved in yeast and plants. We hypothesized that effectors also target conserved cellular processes that are required for yeast growth only under stress conditions. We therefore examined phenotypes induced by expression of Pst effectors in yeast grown in the presence of various stressors. Out of 29 effectors tested, five (HopX1, HopG1, HopT1-1, HopH1 and AvrPtoB) displayed growth inhibition phenotypes only in combination with stress conditions. Viability assays revealed that the HopX1 effector caused loss of cell viability under prolonged osmotic stress. Using transcription reporters, we found that HopX1 attenuated the activation of the high osmolarity glycerol (HOG) mitogen-activated protein kinase (MAPK) pathway, which is responsible for yeast survival under osmotic stress, while other MAPK pathways were mildly affected by HopX1. Interestingly, HopX1-mediated phenotypes in yeast were dependent on the putative transglutaminase catalytic triad of the effector. This study enlarges the pool of phenotypes available for the functional analysis of Pst type III effectors in yeast, and exemplifies how analysis of phenotypes detected in yeast under stress conditions can lead to the identification of eukaryotic cellular processes affected by bacterial effectors.
authors.affiliation: Department of Molecular Biology and Ecology of Plants, Tel Aviv University, 69978 Tel Aviv, Israel.
authors.forename: Dor
authors.initials: D
authors.lastname: Salomon
authors.affiliation: Department of Molecular Biology and Ecology of Plants, Tel Aviv University, 69978 Tel Aviv, Israel.
authors.forename: Eran
authors.initials: E
authors.lastname: Bosis
authors.affiliation: Department of Biochemistry and Molecular Biology, Tel Aviv University, 69978 Tel Aviv, Israel.|Department of Molecular Biology and Ecology of Plants, Tel Aviv University, 69978 Tel Aviv, Israel.
authors.forename: Daniel
authors.initials: D
authors.lastname: Dar
authors.affiliation: Department of Biochemistry and Molecular Biology, Tel Aviv University, 69978 Tel Aviv, Israel.
authors.forename: Iftach
authors.initials: I
authors.lastname: Nachman
authors.affiliation: Department of Molecular Biology and Ecology of Plants, Tel Aviv University, 69978 Tel Aviv, Israel.
authors.forename: Guido
authors.initials: G
authors.lastname: Sessa
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D005990:Glycerol / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics* / Q000378:metabolism; D009994:Osmolar Concentration; D010935:Plant Diseases / Q000382:microbiology; D044224:Pseudomonas syringae / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D029701:Saccharomyces cerevisiae Proteins; D020928:Mitogen-Activated Protein Kinases; D005990:Glycerol