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J Proteome Res ; 13(8): 3635-44, 2014 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-24965097

RESUMO

A proteogenomic analysis is presented for Venturia pirina, a fungus that causes scab disease on European pear (Pyrus communis). V. pirina is host-specific, and the infection is thought to be mediated by secreted effector proteins. Currently, only 36 V. pirina proteins are catalogued in GenBank, and the genome sequence is not publicly available. To identify putative effectors, V. pirina was grown in vitro on and in cellophane sheets mimicking its growth in infected leaves. Secreted extracts were analyzed by tandem mass spectrometry, and the data (ProteomeXchange identifier PXD000710) was queried against a protein database generated by combining in silico predicted transcripts with six frame translations of a whole genome sequence of V. pirina (GenBank Accession JEMP00000000 ). We identified 1088 distinct V. pirina protein groups (FDR 1%) including 1085 detected for the first time. Thirty novel (not in silico predicted) proteins were found, of which 14 were identified as potential effectors based on characteristic features of fungal effector protein sequences. We also used evidence from semitryptic peptides at the protein N-terminus to corroborate in silico signal peptide predictions for 22 proteins, including several potential effectors. The analysis highlights the utility of proteogenomics in the study of secreted effectors.


Assuntos
Ascomicetos/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Proteoma/genética , Pyrus/microbiologia , Ascomicetos/metabolismo , Cromatografia Líquida , Bases de Dados de Proteínas , Genômica/métodos , Folhas de Planta/genética , Folhas de Planta/microbiologia , Proteômica/métodos , Espectrometria de Massas em Tandem
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