title: Loss of Dicer in Sertoli cells has a major impact on the testicular proteome of mice.
abstract: Sertoli cells (SCs) are the central, essential coordinators of spermatogenesis, without which germ cell development cannot occur. We previously showed that Dicer, an RNaseIII endonuclease required for microRNA (miRNA) biogenesis, is absolutely essential for Sertoli cells to mature, survive, and ultimately sustain germ cell development. Here, using isotope-coded protein labeling, a technique for protein relative quantification by mass spectrometry, we investigated the impact of Sertoli cell-Dicer and subsequent miRNA loss on the testicular proteome. We found that, a large proportion of proteins (50 out of 130) are up-regulated by more that 1.3-fold in testes lacking Sertoli cell-Dicer, yet that this protein up-regulation is mild, never exceeding a 2-fold change, and is not preceeded by alterations of the corresponding mRNAs. Of note, the expression levels of six proteins of interest were further validated using the Absolute Quantification (AQUA) peptide technology. Furthermore, through 3'UTR luciferase assays we identified one up-regulated protein, SOD-1, a Cu/Zn superoxide dismutase whose overexpression has been linked to enhanced cell death through apoptosis, as a likely direct target of three Sertoli cell-expressed miRNAs, miR-125a-3p, miR-872 and miR-24. Altogether, our study, which is one of the few in vivo analyses of miRNA effects on protein output, suggests that, at least in our system, miRNAs play a significant role in translation control.
authors.affiliation: Department of Genetic Medicine and Development, University of Geneva Medical School, Geneva, Switzerland.
authors.forename: Marilena D
authors.initials: MD
authors.lastname: Papaioannou
authors.forename: Mélanie
authors.initials: M
authors.lastname: Lagarrigue
authors.forename: Charles E
authors.initials: CE
authors.lastname: Vejnar
authors.forename: Antoine D
authors.initials: AD
authors.lastname: Rolland
authors.forename: Françoise
authors.initials: F
authors.lastname: Kühne
authors.forename: Florence
authors.initials: F
authors.lastname: Aubry
authors.forename: Olivier
authors.initials: O
authors.lastname: Schaad
authors.forename: Alexandre
authors.initials: A
authors.lastname: Fort
authors.forename: Patrick
authors.initials: P
authors.lastname: Descombes
authors.forename: Marguerite
authors.initials: M
authors.lastname: Neerman-Arbez
authors.forename: Florian
authors.initials: F
authors.lastname: Guillou
authors.forename: Evgeny M
authors.initials: EM
authors.lastname: Zdobnov
authors.forename: Charles
authors.initials: C
authors.lastname: Pineau
authors.forename: Serge
authors.initials: S
authors.lastname: Nef
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020543:Proteome / Q000378:metabolism*; D034622:RNA Interference; D043244:Ribonuclease III / Q000172:deficiency* / Q000235:genetics; D017384:Sequence Deletion; D012708:Sertoli Cells / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D000072105:Superoxide Dismutase-1; D053719:Tandem Mass Spectrometry; D013737:Testis / Q000378:metabolism* / Q000473:pathology; D014158:Transcription, Genetic; D015854:Up-Regulation
chemical_list: D020413:3' Untranslated Regions; D035683:MicroRNAs; D020543:Proteome; D049409:Luciferases, Firefly; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D043244:Ribonuclease III
references.citation: Mol Cell Proteomics. 2007 Dec;6(12):2139-49
references.pmid: 17848587
references.citation: Nat Rev Mol Cell Biol. 2009 Feb;10(2):126-39
references.pmid: 19165215
references.citation: Nat Rev Genet. 2002 Oct;3(10):790-801
references.pmid: 12360237
references.citation: Proc Natl Acad Sci U S A. 2008 Feb 5;105(5):1608-13
references.pmid: 18227514
references.citation: EMBO J. 1995 Oct 16;14(20):4985-93
references.pmid: 7588627
references.citation: Nat Genet. 2007 Oct;39(10):1278-84
references.pmid: 17893677
references.citation: J Androl. 2010 Jan-Feb;31(1):26-33
references.pmid: 19875496
references.citation: Cancer Res. 2008 Jul 15;68(14):5540-5
references.pmid: 18632605
references.citation: Nature. 2008 Sep 4;455(7209):58-63
references.pmid: 18668040
references.citation: Nat Rev Genet. 2008 Feb;9(2):102-14
references.pmid: 18197166
references.citation: Science. 2006 Apr 7;312(5770):75-9
references.pmid: 16484454
references.citation: PLoS One. 2008 Mar 05;3(3):e1738
references.pmid: 18320056
references.citation: Science. 2000 Feb 25;287(5457):1489-93
references.pmid: 10688798
references.citation: Nature. 2005 Feb 17;433(7027):769-73
references.pmid: 15685193
references.citation: J Endocrinol. 1989 Sep;122(3):757-62
references.pmid: 2509618
references.citation: Proteomics. 2006 Apr;6(8):2407-21
references.pmid: 16548065
references.citation: Nat Genet. 2004 Jun;36(6):653-9
references.pmid: 15156143
references.citation: Genome Res. 2009 Nov;19(11):2009-20
references.pmid: 19767416
references.citation: Genome Biol. 2002 Jun 18;3(7):RESEARCH0034
references.pmid: 12184808
references.citation: Mol Cell. 2007 Jul 6;27(1):91-105
references.pmid: 17612493
references.citation: Proteomics. 2009 Mar;9(5):1374-84
references.pmid: 19253296
references.citation: Biochem Biophys Res Commun. 2001 Oct 5;287(4):1003-8
references.pmid: 11573964
references.citation: Nature. 2005 Aug 18;436(7053):1030-4
references.pmid: 16107850
references.citation: Methods Enzymol. 1993;225:84-113
references.pmid: 8231890
references.citation: J Biol Chem. 1995 Jan 6;270(1):236-43
references.pmid: 7814380
references.citation: Nucleic Acids Res. 2008 Jan;36(Database issue):D773-9
references.pmid: 18086701
references.citation: Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):6940-5
references.pmid: 12771378
references.citation: Int Rev Cytol. 1993;147:25-96
references.pmid: 8225836
references.citation: Cancer Res. 2009 Apr 15;69(8):3245-8
references.pmid: 19351814
references.citation: J Androl. 2008 Sep-Oct;29(5):488-98
references.pmid: 18567643
references.citation: Biol Reprod. 2008 Oct;79(4):696-703
references.pmid: 18633141
references.citation: Baillieres Clin Endocrinol Metab. 1992 Apr;6(2):273-311
references.pmid: 1616446
references.citation: Science. 2007 Dec 21;318(5858):1931-4
references.pmid: 18048652
references.citation: J Biol Chem. 1999 Feb 26;274(9):5909-18
references.pmid: 10026215
references.citation: Semin Cell Dev Biol. 1998 Aug;9(4):483-9
references.pmid: 9813196
references.citation: Nucleic Acids Res. 2003 Jul 15;31(14):e73
references.pmid: 12853650
references.citation: Cell. 2005 Aug 26;122(4):553-63
references.pmid: 16122423
references.citation: Nucleic Acids Res. 2006;34(16):e107
references.pmid: 16945957
references.citation: Nature. 2008 Sep 4;455(7209):64-71
references.pmid: 18668037
references.citation: Proteomics. 2005 Jan;5(1):4-15
references.pmid: 15602776
references.citation: Dev Biol. 2009 Feb 1;326(1):250-9
references.pmid: 19071104