title: Shotgun proteomics of Aspergillus niger microsomes upon D-xylose induction.
abstract: Protein secretion plays an eminent role in cell maintenance and adaptation to the extracellular environment of microorganisms. Although protein secretion is an extremely efficient process in filamentous fungi, the mechanisms underlying protein secretion have remained largely uncharacterized in these organisms. In this study, we analyzed the effects of the d-xylose induction of cellulase and hemicellulase enzyme secretion on the protein composition of secretory organelles in Aspergillus niger. We aimed to systematically identify the components involved in the secretion of these enzymes via mass spectrometry of enriched subcellular microsomal fractions. Under each condition, fractions enriched for secretory organelles were processed for tandem mass spectrometry, resulting in the identification of peptides that originate from 1,081 proteins, 254 of which-many of them hypothetical proteins-were predicted to play direct roles in the secretory pathway. d-Xylose induction led to an increase in specific small GTPases known to be associated with polarized growth, exocytosis, and endocytosis. Moreover, the endoplasmic-reticulum-associated degradation (ERAD) components Cdc48 and all 14 of the 20S proteasomal subunits were recruited to the secretory organelles. In conclusion, induction of extracellular enzymes results in specific changes in the secretory subproteome of A. niger, and the most prominent change found in this study was the recruitment of the 20S proteasomal subunits to the secretory organelles.
authors.affiliation: Fungal Systems Biology, Laboratory of Systems and Synthetic Biology, Wageningen University, Dreijenplein 10, NL-6708 HB Wageningen, Netherlands.
authors.forename: José Miguel P
authors.initials: JM
authors.lastname: Ferreira de Oliveira
authors.forename: Mark W J
authors.initials: MW
authors.lastname: van Passel
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Schaap
authors.forename: Leo H
authors.initials: LH
authors.lastname: de Graaff
mesh_terms: D001234:Aspergillus niger / Q000235:genetics / Q000254:growth & development / Q000378:metabolism* / Q000648:ultrastructure; D002480:Cellulase / Q000096:biosynthesis; D002853:Chromatography, Liquid; D004721:Endoplasmic Reticulum / Q000201:enzymology / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D006026:Glycoside Hydrolases / Q000378:metabolism; D013058:Mass Spectrometry; D008861:Microsomes / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry; D014994:Xylose / Q000494:pharmacology*
chemical_list: D014994:Xylose; D006026:Glycoside Hydrolases; C023305:hemicellulase; D002480:Cellulase; D046988:Proteasome Endopeptidase Complex; D020558:GTP Phosphohydrolases
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