RT Journal Article SR Electronic T1 The crystal structure of SnTox3 from the necrotrophic fungus Parastagonospora nodorum reveals a unique effector fold and insights into Kex2 protease processing of fungal effectors JF bioRxiv FD Cold Spring Harbor Laboratory SP 2020.05.27.120113 DO 10.1101/2020.05.27.120113 A1 Megan A. Outram A1 Yi-Chang Sung A1 Daniel Yu A1 Bayantes Dagvadorj A1 Sharmin A. Rima A1 David A. Jones A1 Daniel J. Ericsson A1 Jana Sperschneider A1 Peter S. Solomon A1 Bostjan Kobe A1 Simon J. Williams YR 2020 UL http://biorxiv.org/content/early/2020/05/27/2020.05.27.120113.abstract AB Plant pathogens cause disease through secreted effector proteins, which act to modulate host physiology and promote infection. Typically, the sequences of effectors provide little functional information and further targeted experimentation is required. Here, we utilised a structure/function approach to study SnTox3, an effector from the necrotrophic fungal pathogen Parastagonospora nodorum, which causes cell death in wheat-lines carrying the sensitivity gene Snn3.We developed a workflow for the production of SnTox3 in a heterologous host that enabled crystal structure determination. We show this approach can be successfully applied to effectors from other pathogenic fungi. Complementing this, an in-silico study uncovered the prevalence of an expanded subclass of effectors from fungi.The β-barrel fold of SnTox3 is a novel fold among fungal effectors. We demonstrate that SnTox3 is a pre-pro-protein and that the protease Kex2 removes the pro-domain. Our in-silico studies suggest that Kex2-processed pro-domain (designated here as K2PP) effectors are common in fungi, and we demonstrate this experimentally for effectors from Fusarium oxysporum f sp. lycopersici.We propose that K2PP effectors are highly prevalent among fungal effectors. The identification and classification of K2PP effectors has broad implications for the approaches used to study their function in fungal virulence.Competing Interest StatementThe authors have declared no competing interest.