@article{mbs:/content/journal/micro/10.1099/mic.0.29186-0, author = "Rodríguez-Escudero, Isabel and Rotger, Rafael and Cid, Víctor J. and Molina, María", title = "Inhibition of Cdc42-dependent signalling in Saccharomyces cerevisiae by phosphatase-dead SigD/SopB from Salmonella typhimurium", journal= "Microbiology", year = "2006", volume = "152", number = "11", pages = "3437-3452", doi = "https://doi.org/10.1099/mic.0.29186-0", url = "https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.29186-0", publisher = "Microbiology Society", issn = "1465-2080", type = "Journal Article", keywords = "GEF, GDP–GTP exchange factor", keywords = "TTSS, type III secretion system", keywords = "DAPI, 4,6-diamidino-2-phenylindole", keywords = "PtdIns 4,5-P2, phosphatidylinositol 4,5-bisphosphate", keywords = "MAPK, mitogen-activated protein kinase", keywords = "GAP, GTPase-activating protein", keywords = "PtdIns 3,4,5-P3, phosphatidylinositol-3,4,5-trisphosphate", keywords = "GST, glutathione S-transferase", abstract = "Heterologous expression of bacterial virulence factors in Saccharomyces cerevisiae is a feasible approach to study their molecular function. The authors have previously reported that the Salmonella typhimurium SigD protein, a phosphatidylinositol phosphatase involved in invasion of the host cell, inhibits yeast growth, presumably by depleting an essential pool of phosphatidylinositol 4,5-bisphosphate, and also that a catalytically inactive version, SigDR468A, was able to arrest growth by a different mechanism that involved disruption of the actin cytoskeleton. This paper describes marked differences between the phenotypes elicited by expression of SigD and SigDR468A in yeast. First, expression of SigDR468A caused accumulation of large unbudded cells and loss of septin organization, while SigD expression caused none of these effects. Second, growth inhibition by SigDR468A was mediated by a cell cycle arrest in G2 dependent on the Swe1 morphogenetic checkpoint, but SigD-induced growth inhibition was cell cycle independent. And third, SigD caused strong activation of the yeast MAP kinase Slt2, whereas SigDR468A rather inactivated another MAP kinase, Kss1. In a screen for suppressors of SigDR468A-induced growth arrest by overexpression of a yeast cDNA library, the Cdc42 GTPase was isolated. Furthermore, SigDR468A was co-purified with Cdc42 from yeast lysates. It is concluded that the Salmonella SigD protein deprived of its phosphatase activity is able to disrupt yeast morphogenesis by interfering with Cdc42 function, opening the possibility that the SigD N-terminal region might directly modulate small GTPases from the host during infection.", }