title: A Botrytis cinerea emopamil binding domain protein, required for full virulence, belongs to a eukaryotic superfamily which has expanded in euascomycetes.
abstract: A previous transcriptomic analysis of 3,032 fungal genes identified the Botrytis cinerea PIE3 (BcPIE3) gene to be up-regulated early in planta (A. Gioti, A. Simon, P. Le Pêcheur, C. Giraud, J. M. Pradier, M. Viaud, and C. Levis, J. Mol. Biol. 358:372-386, 2006). In the present study, BcPIE3 was disrupted in order to determine its implication in pathogenicity. BcPIE3 was shown to be a virulence factor, since the DeltaBcPIE3 mutant was blocked during the colonization of tomato and bean leaves, giving lesions reduced in size by at least 74%. Within the emopamil binding domain (EBD), BcPIE3 shows significant structural similarities to mammalian emopamil binding proteins (EBPs). Mammalian EBPs function as sterol isomerases, but an analysis of the sterol content and the results of growth inhibition experiments with the DeltaBcPIE3 strain indicated that BcPIE3 is dispensable for ergosterol biosynthesis. The systematic identification of EBD-containing proteins included in public databases showed that these proteins constitute a protein superfamily present only in eukaryotes. Phylogenetic analysis showed that the ancestral EBD-encoding gene was duplicated in the common ancestor of animals and fungi after the split from plants. Finally, we present evidence that the EBP phylogenetic clade of this superfamily has further expanded exclusively in euascomycetes, especially in B. cinerea, which contains three copies of the EBP gene.
authors.affiliation: UMR1290 BIOGER-CPP, INRA, Route de St-Cyr, 78026 Versailles, France. natassa_g_2000@yahoo.com
authors.forename: A
authors.initials: A
authors.lastname: Gioti
authors.forename: J M
authors.initials: JM
authors.lastname: Pradier
authors.forename: E
authors.initials: E
authors.lastname: Fournier
authors.forename: P
authors.initials: P
authors.lastname: Le Pêcheur
authors.forename: C
authors.initials: C
authors.lastname: Giraud
authors.forename: D
authors.initials: D
authors.lastname: Debieu
authors.forename: J
authors.initials: J
authors.lastname: Bach
authors.forename: P
authors.initials: P
authors.lastname: Leroux
authors.forename: C
authors.initials: C
authors.lastname: Levis
mesh_terms: D000595:Amino Acid Sequence; D001203:Ascomycota; D020171:Botrytis / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000502:physiology*; D018551:Solanum lycopersicum / Q000235:genetics / Q000378:metabolism / Q000382:microbiology*; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism / Q000382:microbiology*; D013172:Spores, Fungal / Q000502:physiology; D013251:Steroid Isomerases / Q000235:genetics / Q000378:metabolism*; D013261:Sterols / Q000494:pharmacology; D014774:Virulence
chemical_list: D005656:Fungal Proteins; D013261:Sterols; D013251:Steroid Isomerases