title: MicroRNA-322 (miR-322) and its target protein Tob2 modulate Osterix (Osx) mRNA stability.
abstract: Osteogenesis depends on a coordinated network of signals and transcription factors such as Runx2 and Osterix. Recent evidence indicates that microRNAs (miRNAs) act as important post-transcriptional regulators in a large number of processes, including osteoblast differentiation. In this study, we performed miRNA expression profiling and identified miR-322, a BMP-2-down-regulated miRNA, as a regulator of osteoblast differentiation. We report miR-322 gain- and loss-of-function experiments in C2C12 and MC3T3-E1 cells and primary cultures of murine bone marrow-derived mesenchymal stem cells. We demonstrate that overexpression of miR-322 enhances BMP-2 response, increasing the expression of Osx and other osteogenic genes. Furthermore, we identify Tob2 as a target of miR-322, and we characterize the specific Tob2 3'-UTR sequence bound by miR-322 by reporter assays. We demonstrate that Tob2 is a negative regulator of osteogenesis that binds and mediates degradation of Osx mRNA. Our results demonstrate a new molecular mechanism controlling osteogenesis through the specific miR-322/Tob2 regulation of specific target mRNAs. This regulatory circuit provides a clear example of a complex miRNA-transcription factor network for fine-tuning the osteoblast differentiation program.
authors.affiliation: Departament de Ciències Fisiològiques II, Universitat de Barcelona, and L'Institut d'Investigació Biomèdica de Bellvitge (IDIBELL), E-08907 L'Hospitalet de Llobregat, Barcelona, Spain.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Gámez
authors.affiliation: Departament de Ciències Fisiològiques II, Universitat de Barcelona, and L'Institut d'Investigació Biomèdica de Bellvitge (IDIBELL), E-08907 L'Hospitalet de Llobregat, Barcelona, Spain.
authors.forename: Edgardo
authors.initials: E
authors.lastname: Rodríguez-Carballo
authors.affiliation: Departament de Ciències Fisiològiques II, Universitat de Barcelona, and L'Institut d'Investigació Biomèdica de Bellvitge (IDIBELL), E-08907 L'Hospitalet de Llobregat, Barcelona, Spain.
authors.forename: Ramon
authors.initials: R
authors.lastname: Bartrons
authors.affiliation: Departament de Ciències Fisiològiques II, Universitat de Barcelona, and L'Institut d'Investigació Biomèdica de Bellvitge (IDIBELL), E-08907 L'Hospitalet de Llobregat, Barcelona, Spain.
authors.forename: José Luis
authors.initials: JL
authors.lastname: Rosa
authors.affiliation: Departament de Ciències Fisiològiques II, Universitat de Barcelona, and L'Institut d'Investigació Biomèdica de Bellvitge (IDIBELL), E-08907 L'Hospitalet de Llobregat, Barcelona, Spain. Electronic address: fventura@ub.edu.
authors.forename: Francesc
authors.initials: F
authors.lastname: Ventura
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000166:cytology*; D010012:Osteogenesis / Q000502:physiology*; D020871:RNA Stability; D012333:RNA, Messenger / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D000076182:Sp7 Transcription Factor; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; C582672:MIRN322 microRNA, mouse; D035683:MicroRNAs; D012333:RNA, Messenger; D000076182:Sp7 Transcription Factor; C479466:Sp7 protein, mouse; C485732:Tob2 protein, mouse; D014157:Transcription Factors
keywords: Bone; Bone Morphogenetic Protein (BMP); MicroRNA; Osteoblast Differentiation; Osteoblasts; Osterix; Polyadenylation; Tob2; mRNA Decay; mRNA Degradation
references.citation: Growth Factors. 2004 Dec;22(4):233-41
references.pmid: 15621726
references.citation: Nucleic Acids Res. 2012 Jul;40(Web Server issue):W498-504
references.pmid: 22649059
references.citation: EMBO J. 2006 Oct 18;25(20):4865-76
references.pmid: 17024188
references.citation: Science. 2008 Mar 28;319(5871):1785-6
references.pmid: 18369135
references.citation: Nat Rev Genet. 2012 Mar 06;13(4):246-59
references.pmid: 22392217
references.citation: EMBO J. 2011 Apr 6;30(7):1311-23
references.pmid: 21336257
references.citation: Mol Cell Biol. 2007 Nov;27(22):7791-801
references.pmid: 17785442
references.citation: J Cell Biochem. 1999 Apr 1;73(1):114-25
references.pmid: 10088730
references.citation: J Clin Invest. 2009 Dec;119(12):3666-77
references.pmid: 19920351
references.citation: Mol Cell. 2010 Aug 13;39(3):373-84
references.pmid: 20705240
references.citation: J Cell Physiol. 2010 Jan;222(1):66-72
references.pmid: 19746446
references.citation: Trends Genet. 2003 Aug;19(8):458-66
references.pmid: 12902164
references.citation: J Cell Biol. 1994 Dec;127(6 Pt 1):1755-66
references.pmid: 7798324
references.citation: J Biol Chem. 2008 Feb 15;283(7):3816-26
references.pmid: 18056716
references.citation: Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6653-8
references.pmid: 15100414
references.citation: J Biol Chem. 2004 Feb 13;279(7):5397-404
references.pmid: 14645243
references.citation: Cell. 1997 May 30;89(5):747-54
references.pmid: 9182762
references.citation: J Bone Miner Res. 2007 Aug;22(8):1217-23
references.pmid: 17451368
references.citation: Mol Biol Cell. 2010 Jul 1;21(13):2138-49
references.pmid: 20462953
references.citation: Genes Dev. 2007 Dec 1;21(23):3135-48
references.pmid: 18056425
references.citation: Genes Dev. 2005 Dec 1;19(23):2783-810
references.pmid: 16322555
references.citation: Biochem Biophys Res Commun. 2009 May 22;383(1):125-9
references.pmid: 19341710
references.citation: PLoS Biol. 2004 Nov;2(11):e363
references.pmid: 15502875
references.citation: J Biol Chem. 2010 Aug 13;285(33):25221-31
references.pmid: 20551325
references.citation: Mol Cell. 2007 Jul 6;27(1):91-105
references.pmid: 17612493
references.citation: J Biol Chem. 2002 Feb 1;277(5):3176-85
references.pmid: 11700304
references.citation: J Biol Chem. 2012 Mar 2;287(10):7503-11
references.pmid: 22253433
references.citation: Dev Biol. 2010 Apr 1;340(1):10-21
references.pmid: 20079730
references.citation: J Bone Miner Res. 2011 Apr;26(4):718-29
references.pmid: 20878775
references.citation: Trends Biochem Sci. 2009 Dec;34(12):640-7
references.pmid: 19828319
references.citation: Proc Natl Acad Sci U S A. 2009 Dec 8;106(49):20794-9
references.pmid: 19933329
references.citation: Int J Biochem Cell Biol. 2008;40(3):383-408
references.pmid: 18061509
references.citation: Eur J Endocrinol. 2012 Mar;166(3):359-71
references.pmid: 22084154
references.citation: Mol Cell Biol. 2012 Mar;32(6):1089-98
references.pmid: 22252318
references.citation: Ann Surg Oncol. 2009 Jun;16(6):1704-10
references.pmid: 19326169
references.citation: J Am Soc Nephrol. 2011 Aug;22(8):1462-74
references.pmid: 21784902
references.citation: Genes Dev. 2011 Oct 15;25(20):2173-86
references.pmid: 22012620
references.citation: Cell. 2002 Jan 11;108(1):17-29
references.pmid: 11792318
references.citation: FEBS Lett. 2012 Jul 4;586(14):1906-12
references.pmid: 22306316
references.citation: Nucleic Acids Res. 2008 May;36(8):2690-9
references.pmid: 18353861
references.citation: Nature. 2008 Jul 3;454(7200):56-61
references.pmid: 18548003
references.citation: Biochem Biophys Res Commun. 2005 Mar 18;328(3):651-7
references.pmid: 15694398
references.citation: Mol Endocrinol. 2011 Jun;25(6):1006-17
references.pmid: 21436263
references.citation: J Biol Chem. 2010 Oct 15;285(42):31985-94
references.pmid: 20682789
references.citation: Proc Natl Acad Sci U S A. 2011 Jun 14;108(24):9863-8
references.pmid: 21628588
references.citation: Proc Natl Acad Sci U S A. 2008 Sep 16;105(37):13906-11
references.pmid: 18784367
references.citation: Cell. 2000 Dec 22;103(7):1085-97
references.pmid: 11163184
references.citation: Cell. 2004 Jan 23;116(2):281-97
references.pmid: 14744438
references.citation: J Bone Miner Res. 2008 Feb;23(2):287-95
references.pmid: 18197755
references.citation: Curr Opin Cell Biol. 2009 Jun;21(3):461-9
references.pmid: 19278845
references.citation: Am J Hum Genet. 2005 Nov;77(5):807-23
references.pmid: 16252240
references.citation: Annu Rev Genomics Hum Genet. 2008;9:183-96
references.pmid: 18767962
references.citation: Nat Rev Endocrinol. 2012 Jan 31;8(4):212-27
references.pmid: 22290358