title: Compatibility in the Ustilago maydis-maize interaction requires inhibition of host cysteine proteases by the fungal effector Pit2.
abstract: The basidiomycete Ustilago maydis causes smut disease in maize, with large plant tumors being formed as the most prominent disease symptoms. During all steps of infection, U. maydis depends on a biotrophic interaction, which requires an efficient suppression of plant immunity. In a previous study, we identified the secreted effector protein Pit2, which is essential for maintenance of biotrophy and induction of tumors. Deletion mutants for pit2 successfully penetrate host cells but elicit various defense responses, which stops further fungal proliferation. We now show that Pit2 functions as an inhibitor of a set of apoplastic maize cysteine proteases, whose activity is directly linked with salicylic-acid-associated plant defenses. Consequently, protease inhibition by Pit2 is required for U. maydis virulence. Sequence comparisons with Pit2 orthologs from related smut fungi identified a conserved sequence motif. Mutation of this sequence caused loss of Pit2 function. Consequently, expression of the mutated protein in U. maydis could not restore virulence of the pit2 deletion mutant, indicating that the protease inhibition by Pit2 is essential for fungal virulence. Moreover, synthetic peptides of the conserved sequence motif showed full activity as protease inhibitor, which identifies this domain as a new, minimal protease inhibitor domain in plant-pathogenic fungi.
authors: Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.
authors: André N
authors: AN
authors: Mueller
authors: Sebastian
authors: S
authors: Ziemann
authors: Steffi
authors: S
authors: Treitschke
authors: Daniela
authors: D
authors: Aßmann
authors: Gunther
authors: G
authors: Doehlemann
mesh_terms: D000595:Amino Acid Sequence; D057056:Cysteine Proteases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D010935:Plant Diseases / Q000235:genetics / Q000276:immunology / Q000382:microbiology*; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D020798:Two-Hybrid System Techniques; D014588:Ustilago / Q000472:pathogenicity / Q000502:physiology*; D014774:Virulence; D003313:Zea mays / Q000201:enzymology* / Q000382:microbiology
chemical_list: D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D010446:Peptide Fragments; D057056:Cysteine Proteases