Protein kinase D2 is an essential regulator of murine myoblast differentiation.

Muscle differentiation is a highly conserved process that occurs through the activation of quiescent satellite cells whose progeny proliferate, differentiate, and fuse to generate new myofibers. A defined pattern of myogenic transcription factors is orchestrated during this process and is regulated...

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Autores principales: Alexander Kleger, Christiane Loebnitz, Ganesh V Pusapati, Milena Armacki, Martin Müller, Stefan Tümpel, Anett Illing, Daniel Hartmann, Cornelia Brunner, Stefan Liebau, Karl L Rudolph, Guido Adler, Thomas Seufferlein
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Publicado: Public Library of Science (PLoS) 2011
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spelling oai:doaj.org-article:2b71aaf2aee84de9b899cc7c44efc6492021-11-18T06:59:44ZProtein kinase D2 is an essential regulator of murine myoblast differentiation.1932-620310.1371/journal.pone.0014599https://doaj.org/article/2b71aaf2aee84de9b899cc7c44efc6492011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21298052/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Muscle differentiation is a highly conserved process that occurs through the activation of quiescent satellite cells whose progeny proliferate, differentiate, and fuse to generate new myofibers. A defined pattern of myogenic transcription factors is orchestrated during this process and is regulated via distinct signaling cascades involving various intracellular signaling pathways, including members of the protein kinase C (PKC) family. The protein kinase D (PKD) isoenzymes PKD1, -2, and -3, are prominent downstream targets of PKCs and phospholipase D in various biological systems including mouse and could hence play a role in muscle differentiation. In the present study, we used a mouse myoblast cell line (C2C12) as an in vitro model to investigate the role of PKDs, in particular PKD2, in muscle stem cell differentiation. We show that C2C12 cells express all PKD isoforms with PKD2 being highly expressed. Furthermore, we demonstrate that PKD2 is specifically phosphorylated/activated during the initiation of mouse myoblast differentiation. Selective inhibition of PKCs or PKDs by pharmacological inhibitors blocked myotube formation. Depletion of PKD2 by shRNAs resulted in a marked inhibition of myoblast cell fusion. PKD2-depleted cells exhibit impaired regulation of muscle development-associated genes while the proliferative capacity remains unaltered. Vice versa forced expression of PKD2 increases myoblast differentiation. These findings were confirmed in primary mouse satellite cells where myotube fusion was also decreased upon inhibition of PKDs. Active PKD2 induced transcriptional activation of myocyte enhancer factor 2D and repression of Pax3 transcriptional activity. In conclusion, we identify PKDs, in particular PKD2, as a major mediator of muscle cell differentiation in vitro and thereby as a potential novel target for the modulation of muscle regeneration.Alexander KlegerChristiane LoebnitzGanesh V PusapatiMilena ArmackiMartin MüllerStefan TümpelAnett IllingDaniel HartmannCornelia BrunnerStefan LiebauKarl L RudolphGuido AdlerThomas SeufferleinPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 1, p e14599 (2011)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Alexander Kleger
Christiane Loebnitz
Ganesh V Pusapati
Milena Armacki
Martin Müller
Stefan Tümpel
Anett Illing
Daniel Hartmann
Cornelia Brunner
Stefan Liebau
Karl L Rudolph
Guido Adler
Thomas Seufferlein
Protein kinase D2 is an essential regulator of murine myoblast differentiation.
description Muscle differentiation is a highly conserved process that occurs through the activation of quiescent satellite cells whose progeny proliferate, differentiate, and fuse to generate new myofibers. A defined pattern of myogenic transcription factors is orchestrated during this process and is regulated via distinct signaling cascades involving various intracellular signaling pathways, including members of the protein kinase C (PKC) family. The protein kinase D (PKD) isoenzymes PKD1, -2, and -3, are prominent downstream targets of PKCs and phospholipase D in various biological systems including mouse and could hence play a role in muscle differentiation. In the present study, we used a mouse myoblast cell line (C2C12) as an in vitro model to investigate the role of PKDs, in particular PKD2, in muscle stem cell differentiation. We show that C2C12 cells express all PKD isoforms with PKD2 being highly expressed. Furthermore, we demonstrate that PKD2 is specifically phosphorylated/activated during the initiation of mouse myoblast differentiation. Selective inhibition of PKCs or PKDs by pharmacological inhibitors blocked myotube formation. Depletion of PKD2 by shRNAs resulted in a marked inhibition of myoblast cell fusion. PKD2-depleted cells exhibit impaired regulation of muscle development-associated genes while the proliferative capacity remains unaltered. Vice versa forced expression of PKD2 increases myoblast differentiation. These findings were confirmed in primary mouse satellite cells where myotube fusion was also decreased upon inhibition of PKDs. Active PKD2 induced transcriptional activation of myocyte enhancer factor 2D and repression of Pax3 transcriptional activity. In conclusion, we identify PKDs, in particular PKD2, as a major mediator of muscle cell differentiation in vitro and thereby as a potential novel target for the modulation of muscle regeneration.
format article
author Alexander Kleger
Christiane Loebnitz
Ganesh V Pusapati
Milena Armacki
Martin Müller
Stefan Tümpel
Anett Illing
Daniel Hartmann
Cornelia Brunner
Stefan Liebau
Karl L Rudolph
Guido Adler
Thomas Seufferlein
author_facet Alexander Kleger
Christiane Loebnitz
Ganesh V Pusapati
Milena Armacki
Martin Müller
Stefan Tümpel
Anett Illing
Daniel Hartmann
Cornelia Brunner
Stefan Liebau
Karl L Rudolph
Guido Adler
Thomas Seufferlein
author_sort Alexander Kleger
title Protein kinase D2 is an essential regulator of murine myoblast differentiation.
title_short Protein kinase D2 is an essential regulator of murine myoblast differentiation.
title_full Protein kinase D2 is an essential regulator of murine myoblast differentiation.
title_fullStr Protein kinase D2 is an essential regulator of murine myoblast differentiation.
title_full_unstemmed Protein kinase D2 is an essential regulator of murine myoblast differentiation.
title_sort protein kinase d2 is an essential regulator of murine myoblast differentiation.
publisher Public Library of Science (PLoS)
publishDate 2011
url https://doaj.org/article/2b71aaf2aee84de9b899cc7c44efc649
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