HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 278, Issue 47, 47119-47128, November 21, 2003

This Article Full Text Full Text (PDF) All Versions of this Article: 278/47/47119    most recent M308889200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by van der Giessen, K. Articles by Gallouzi, I. E. Search for Related Content PubMed PubMed Citation Articles by van der Giessen, K. Articles by Gallouzi, I. E. Social Bookmarking                      What's this?

RNAi-mediated HuR Depletion Leads to the Inhibition of Muscle Cell Differentiation^*

Kate van der Giessen, Sergio Di-Marco, Eveline Clair, and Imed Eddine Gallouzi

From the Department of Biochemistry, McGill University, Montreal, Quebec H3G 1Y6, Canada

The formation of muscle fibers involves the sequential expression of many proteins that regulate key steps during myoblast-to-myotube transition. MyoD, myogenin, and the cyclin-dependent kinase inhibitor p21^cip1 are major players in the initiation and maintenance of the differentiated state of mouse embryonic muscle cells (C2C12). The messenger RNAs encoding these three proteins contain typical AU-rich elements (AREs) in their 3'-untranslated regions (3'-UTRs), which are known to affect the half-life of many short-lived mRNAs. HuR, an RNA-binding protein that regulates both the stability and cellular movement of ARE-containing mRNAs, interacts and stabilizes the p21^cip1 message under UV stress in human RKO colorectal carcinoma cells. Here, by the use of gel shift experiments and immunoprecipitation followed by reverse transcription-PCR analysis, we show that HuR interacts with MyoD, myogenin, and p21^cip1 mRNAs through specific sequences in their 3'-UTRs. To demonstrate the implication of endogenous HuR in myogenesis, we knocked down its expression in myoblasts using RNA interference and observed a significant reduction of HuR expression, associated with complete inhibition of myogenesis. Moreover, the expression of MyoD and myogenin mRNAs, as well as proteins, is significantly reduced in the HuR knockdown C2C12 cells. We were able to completely re-establish the myogenic process of these defective cells by introducing back HuR protein conjugated to a cell-permeable peptide. Finally, HuR accumulates in the cytoplasm during myogenesis. Thus, our results clearly demonstrated that endogenous HuR plays a crucial role in muscle differentiation by regulating the expression and/or the nuclear export of ARE-containing mRNAs that are essential for this process.

Received for publication, August 12, 2003 , and in revised form, August 27, 2003.

^* This work was supported by a Canadian Institutes for Health Research (CIHR) Cancer Consortium Training Grant Fellowship Award (to S. D.) and CIHR Operating Grant Mop-57680 and a Fond de Recherche Scientifique Quebecois "Subvention d'etablissement de jeune chercheur" Frsq 23516-2760 (to I. E. G.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Biochemistry, McGill University, McIntyre Bldg., Rm. 904, 3655 Promenade Sir William Osler, Montreal, Quebec H3G 1Y6, Canada. Tel.: 514-398-4537; Fax: 514-398-7384; E-mail: imed.gallouzi{at}mcgill.ca.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				V. Katsanou, S. Milatos, A. Yiakouvaki, N. Sgantzis, A. Kotsoni, M. Alexiou, V. Harokopos, V. Aidinis, M. Hemberger, and D. L. Kontoyiannis The RNA-Binding Protein Elavl1/HuR Is Essential for Placental Branching Morphogenesis and Embryonic Development Mol. Cell. Biol., May 15, 2009; 29(10): 2762 - 2776. [Abstract] [Full Text] [PDF]

				H. H. Kim, K. Abdelmohsen, A. Lal, R. Pullmann Jr., X. Yang, S. Galban, S. Srikantan, J. L. Martindale, J. Blethrow, K. M. Shokat, et al. Nuclear HuR accumulation through phosphorylation by Cdk1 Genes & Dev., July 1, 2008; 22(13): 1804 - 1815. [Abstract] [Full Text] [PDF]

				Y. Yamaguchi, R. Oohinata, T. Naiki, and K. Irie Stau1 negatively regulates myogenic differentiation in C2C12 cells. Genes Cells, June 1, 2008; 13(6): 583 - 592. [Abstract] [Full Text] [PDF]

				J. V. Chakkalakal, P. Miura, G. Belanger, R. N. Michel, and B. J. Jasmin Modulation of utrophin A mRNA stability in fast versus slow muscles via an AU-rich element and calcineurin signaling Nucleic Acids Res., February 11, 2008; 36(3): 826 - 838. [Abstract] [Full Text] [PDF]

				R. Mazroui, S. Di Marco, E. Clair, C. von Roretz, S. A. Tenenbaum, J. D. Keene, M. Saleh, and I.-E. Gallouzi Caspase-mediated cleavage of HuR in the cytoplasm contributes to pp32/PHAP-I regulation of apoptosis J. Cell Biol., January 10, 2008; 180(1): 113 - 127. [Abstract] [Full Text] [PDF]

				V. Dormoy-Raclet, I. Menard, E. Clair, G. Kurban, R. Mazroui, S. Di Marco, C. von Roretz, A. Pause, and I.-E. Gallouzi The RNA-Binding Protein HuR Promotes Cell Migration and Cell Invasion by Stabilizing the {beta}-actin mRNA in a U-Rich-Element-Dependent Manner Mol. Cell. Biol., August 1, 2007; 27(15): 5365 - 5380. [Abstract] [Full Text] [PDF]

				K. van der Giessen and I.-E. Gallouzi Involvement of Transportin 2-mediated HuR Import in Muscle Cell Differentiation Mol. Biol. Cell, July 1, 2007; 18(7): 2619 - 2629. [Abstract] [Full Text] [PDF]

				X.-L. Li, J. B. Andersen, H. J. Ezelle, G. M. Wilson, and B. A. Hassel Post-transcriptional Regulation of RNase-L Expression Is Mediated by the 3'-Untranslated Region of Its mRNA J. Biol. Chem., March 16, 2007; 282(11): 7950 - 7960. [Abstract] [Full Text] [PDF]

				R. W. Burry and C. L. Smith HuD Distribution Changes in Response to Heat Shock but Not Neurotrophic Stimulation J. Histochem. Cytochem., October 1, 2006; 54(10): 1129 - 1138. [Abstract] [Full Text] [PDF]

				F. Sato, M. Kurokawa, N. Yamauchi, and M.-a. Hattori Gene silencing of myostatin in differentiation of chicken embryonic myoblasts by small interfering RNA Am J Physiol Cell Physiol, September 1, 2006; 291(3): C538 - C545. [Abstract] [Full Text] [PDF]

				D. T. Fritz, S. Jiang, J. Xu, and M. B. Rogers A Polymorphism in a Conserved Posttranscriptional Regulatory Motif Alters Bone Morphogenetic Protein 2 (BMP2) RNA:Protein Interactions Mol. Endocrinol., July 1, 2006; 20(7): 1574 - 1586. [Abstract] [Full Text] [PDF]

				J. G. Wang, M. Collinge, V. Ramgolam, O. Ayalon, X. C. Fan, R. Pardi, and J. R. Bender LFA-1-Dependent HuR Nuclear Export and Cytokine mRNA Stabilization in T Cell Activation J. Immunol., February 15, 2006; 176(4): 2105 - 2113. [Abstract] [Full Text] [PDF]

				Y. Z. Xu, S. Di Marco, I. Gallouzi, M. Rola-Pleszczynski, and D. Radzioch RNA-Binding Protein HuR Is Required for Stabilization of SLC11A1 mRNA and SLC11A1 Protein Expression Mol. Cell. Biol., September 15, 2005; 25(18): 8139 - 8149. [Abstract] [Full Text] [PDF]

				S. Di Marco, R. Mazroui, P. Dallaire, S. Chittur, S. A. Tenenbaum, D. Radzioch, A. Marette, and I.-E. Gallouzi NF-{kappa}B-Mediated MyoD Decay during Muscle Wasting Requires Nitric Oxide Synthase mRNA Stabilization, HuR Protein, and Nitric Oxide Release Mol. Cell. Biol., August 1, 2005; 25(15): 6533 - 6545. [Abstract] [Full Text] [PDF]

				J. Deschenes-Furry, G. Belanger, J. Mwanjewe, J. A. Lunde, R. J. Parks, N. Perrone-Bizzozero, and B. J. Jasmin The RNA-binding Protein HuR Binds to Acetylcholinesterase Transcripts and Regulates Their Expression in Differentiating Skeletal Muscle Cells J. Biol. Chem., July 8, 2005; 280(27): 25361 - 25368. [Abstract] [Full Text] [PDF]

				K. Gantt, J. Cherry, R. Tenney, V. Karschner, and P. H. Pekala An Early Event in Adipogenesis, the Nuclear Selection of the CCAAT Enhancer-binding Protein {beta} (C/EBP{beta}) mRNA by HuR and its Translocation to the Cytosol J. Biol. Chem., July 1, 2005; 280(26): 24768 - 24774. [Abstract] [Full Text] [PDF]

				R. Pullmann Jr., M. Juhaszova, I. L. de Silanes, T. Kawai, K. Mazan-Mamczarz, M. K. Halushka, and M. Gorospe Enhanced Proliferation of Cultured Human Vascular Smooth Muscle Cells Linked to Increased Function of RNA-binding Protein HuR J. Biol. Chem., June 17, 2005; 280(24): 22819 - 22826. [Abstract] [Full Text] [PDF]

				L. Shi, G. Zhao, D. Qiu, W. R. Godfrey, H. Vogel, T. A. Rando, H. Hu, and P. N. Kao NF90 Regulates Cell Cycle Exit and Terminal Myogenic Differentiation by Direct Binding to the 3'-Untranslated Region of MyoD and p21WAF1/CIP1 mRNAs J. Biol. Chem., May 13, 2005; 280(19): 18981 - 18989. [Abstract] [Full Text] [PDF]

				N. Bonafe, M. Gilmore-Hebert, N. L. Folk, M. Azodi, Y. Zhou, and S. K. Chambers Glyceraldehyde-3-Phosphate Dehydrogenase Binds to the AU-Rich 3' Untranslated Region of Colony-Stimulating Factor-1 (CSF-1) Messenger RNA in Human Ovarian Cancer Cells: Possible Role in CSF-1 Posttranscriptional Regulation and Tumor Phenotype Cancer Res., May 1, 2005; 65(9): 3762 - 3771. [Abstract] [Full Text] [PDF]

				M. Yano, H. J. Okano, and H. Okano Involvement of Hu and Heterogeneous Nuclear Ribonucleoprotein K in Neuronal Differentiation through p21 mRNA Post-transcriptional Regulation J. Biol. Chem., April 1, 2005; 280(13): 12690 - 12699. [Abstract] [Full Text] [PDF]

				I. L. de Silanes, M. Zhan, A. Lal, X. Yang, and M. Gorospe Identification of a target RNA motif for RNA-binding protein HuR PNAS, March 2, 2004; 101(9): 2987 - 2992. [Abstract] [Full Text] [PDF]