Loss of Dicer in Sertoli cells has a major impact on the testicular proteome of mice.
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.
Papaioannou
Marilena D
MD
Department of Genetic Medicine and Development, University of Geneva Medical School, Geneva, Switzerland.
Lagarrigue
Mélanie
M
Vejnar
Charles E
CE
Rolland
Antoine D
AD
Kühne
Françoise
F
Aubry
Florence
F
Schaad
Olivier
O
Fort
Alexandre
A
Descombes
Patrick
P
Neerman-Arbez
Marguerite
M
Guillou
Florian
F
Zdobnov
Evgeny M
EM
Pineau
Charles
C
Nef
Serge
S
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
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
Mol Cell Proteomics. 2007 Dec;6(12):2139-49
17848587
Nat Rev Mol Cell Biol. 2009 Feb;10(2):126-39
19165215
Nat Rev Genet. 2002 Oct;3(10):790-801
12360237
Proc Natl Acad Sci U S A. 2008 Feb 5;105(5):1608-13
18227514
EMBO J. 1995 Oct 16;14(20):4985-93
7588627
Nat Genet. 2007 Oct;39(10):1278-84
17893677
J Androl. 2010 Jan-Feb;31(1):26-33
19875496
Cancer Res. 2008 Jul 15;68(14):5540-5
18632605
Nature. 2008 Sep 4;455(7209):58-63
18668040
Nat Rev Genet. 2008 Feb;9(2):102-14
18197166
Science. 2006 Apr 7;312(5770):75-9
16484454
PLoS One. 2008 Mar 05;3(3):e1738
18320056
Science. 2000 Feb 25;287(5457):1489-93
10688798
Nature. 2005 Feb 17;433(7027):769-73
15685193
J Endocrinol. 1989 Sep;122(3):757-62
2509618
Proteomics. 2006 Apr;6(8):2407-21
16548065
Nat Genet. 2004 Jun;36(6):653-9
15156143
Genome Res. 2009 Nov;19(11):2009-20
19767416
Genome Biol. 2002 Jun 18;3(7):RESEARCH0034
12184808
Mol Cell. 2007 Jul 6;27(1):91-105
17612493
Proteomics. 2009 Mar;9(5):1374-84
19253296
Biochem Biophys Res Commun. 2001 Oct 5;287(4):1003-8
11573964
Nature. 2005 Aug 18;436(7053):1030-4
16107850
Methods Enzymol. 1993;225:84-113
8231890
J Biol Chem. 1995 Jan 6;270(1):236-43
7814380
Nucleic Acids Res. 2008 Jan;36(Database issue):D773-9
18086701
Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):6940-5
12771378
Int Rev Cytol. 1993;147:25-96
8225836
Cancer Res. 2009 Apr 15;69(8):3245-8
19351814
J Androl. 2008 Sep-Oct;29(5):488-98
18567643
Biol Reprod. 2008 Oct;79(4):696-703
18633141
Baillieres Clin Endocrinol Metab. 1992 Apr;6(2):273-311
1616446
Science. 2007 Dec 21;318(5858):1931-4
18048652
J Biol Chem. 1999 Feb 26;274(9):5909-18
10026215
Semin Cell Dev Biol. 1998 Aug;9(4):483-9
9813196
Nucleic Acids Res. 2003 Jul 15;31(14):e73
12853650
Cell. 2005 Aug 26;122(4):553-63
16122423
Nucleic Acids Res. 2006;34(16):e107
16945957
Nature. 2008 Sep 4;455(7209):64-71
18668037
Proteomics. 2005 Jan;5(1):4-15
15602776
Dev Biol. 2009 Feb 1;326(1):250-9
19071104