Shotgun proteomics of Aspergillus niger microsomes upon D-xylose induction.
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.
Ferreira de Oliveira
José Miguel P
JM
Fungal Systems Biology, Laboratory of Systems and Synthetic Biology, Wageningen University, Dreijenplein 10, NL-6708 HB Wageningen, Netherlands.
van Passel
Mark W J
MW
Schaap
Peter J
PJ
de Graaff
Leo H
LH
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*
D014994:Xylose; D006026:Glycoside Hydrolases; C023305:hemicellulase; D002480:Cellulase; D046988:Proteasome Endopeptidase Complex; D020558:GTP Phosphohydrolases
Gene. 1994 Aug 5;145(2):171-8
8056328
Fungal Genet Biol. 2009 Mar;46 Suppl 1:S141-52
18824119
Mol Biol Cell. 2008 Apr;19(4):1439-49
18216285
Mol Microbiol. 1998 Jan;27(1):131-42
9466262
J Cell Biol. 1998 Aug 10;142(3):665-81
9700157
Biotechnology (N Y). 1991 Oct;9(10):976-81
1368725
Mol Biol Cell. 2004 Jun;15(6):2907-19
15047867
Appl Environ Microbiol. 2006 May;72(5):3448-57
16672490
J Proteome Res. 2006 Sep;5(9):2339-47
16944946
Eukaryot Cell. 2006 Dec;5(12):1990-2000
17012537
J Mol Evol. 1998 Nov;47(5):531-43
9797403
Antonie Van Leeuwenhoek. 1987;53(5):349-52
3120641
BMC Genomics. 2007 Jun 11;8:158
17561995
J Cell Biol. 2001 Feb 5;152(3):503-18
11157978
Nat Cell Biol. 2006 May;8(5):485-91
16622418
Cell. 2006 Apr 7;125(1):173-86
16615898
Anal Biochem. 1976 May 7;72:248-54
942051
J Biol Chem. 1994 Nov 4;269(44):27494-502
7961664
Cell. 2006 Dec 15;127(6):1265-81
17174899
Mol Biol Cell. 2009 Oct;20(20):4335-47
19692566
Biotechnol Adv. 2009 Jan-Feb;27(1):53-75
18840517
Curr Opin Plant Biol. 2007 Dec;10(6):594-9
17913569
Proc Int Conf Intell Syst Mol Biol. 1998;6:175-82
9783223
Proc Natl Acad Sci U S A. 2008 Mar 18;105(11):4387-92
18332432
Plant Cell. 2008 Jul;20(7):1948-63
18660430
Can J Microbiol. 1998 Oct;44(10):945-53
9933913
Nat Biotechnol. 2007 Feb;25(2):221-31
17259976
Science. 1998 Jun 12;280(5370):1763-6
9624056
Mol Biol Cell. 1993 Sep;4(9):931-9
8257795
Appl Microbiol Biotechnol. 2008 Oct;80(5):917-24
18704394
J Cell Sci. 2009 Jan 1;122(Pt 1):25-35
19050045
Fungal Genet Biol. 2009 Mar;46 Suppl 1:S153-S160
19618504
Mol Biol Cell. 1999 Nov;10(11):3643-59
10564262
Mol Genet Genomics. 2001 Oct;266(2):246-53
11683266
J Biol Chem. 1996 Oct 25;271(43):26939-46
8900179
J Biol Chem. 2006 Mar 17;281(11):7595-604
16407213
Appl Environ Microbiol. 2009 Apr;75(8):2414-22
19233957
Yeast. 2005 Feb;22(3):219-39
15704212
Adv Genet. 1953;5:141-238
13040135
Gene. 1997 Dec 19;204(1-2):171-6
9434181
Nucleic Acids Res. 2001 May 1;29(9):e45
11328886
Appl Environ Microbiol. 2009 Apr;75(7):2212-20
19168657
Bioinformatics. 2009 May 1;25(9):1189-91
19151095
Microbiology (Reading). 2004 May;150(Pt 5):1367-1375
15133098
Glycobiology. 2000 Oct;10(10):983-91
11030744
Circ Res. 2003 May 16;92(9):962-8
12750306
Fungal Genet Biol. 2004 Jan;41(1):13-22
14643255
Biochim Biophys Acta. 2006 Feb;1762(2):140-7
16054341
Proteomics. 2005 Aug;5(12):3153-61
16035112
Genome Biol. 2007;8(9):R182
17784953
Adv Biochem Eng Biotechnol. 2008;111:99-147
18253709
Mol Microbiol. 2001 Jul;41(2):513-25
11489135
Cell. 2007 Jun 29;129(7):1325-36
17604721
Appl Microbiol Biotechnol. 1997 Jan;47(1):1-11
9035405
J Biol Chem. 1997 Dec 19;272(51):32513-20
9405463
J Biol Chem. 2009 Oct 30;284(44):30328-38
19723633
Oncogene. 2000 Mar 9;19(11):1428-36
10723134
BMC Genomics. 2009 Jan 23;10:44
19166577
Traffic. 2009 Jan;10(1):57-75
19000168
J Proteome Res. 2004 Sep-Oct;3(5):958-64
15473683
Med Mycol. 2006 Sep;44 Suppl 1:S17-21
17050415
Nucleic Acids Res. 1994 Nov 11;22(22):4673-80
7984417