Yap is a novel regulator of C2C12 myogenesis

Kevin I Watt, Robert Judson, Paul Medlow, Kenneth Reid, Tobias B Kurth, Jatin G Burniston, Aivaras Ratkevicius, Cosimo De Bari, Henning Wackerhage

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Abstract

The expression, regulation and function of mammalian Hippo pathway members in skeletal muscle is largely unknown. The aim of this study was thus to test the hypothesis that core members of the mammalian Hippo pathway are expressed in skeletal muscle and that the transcriptional co-factor Yap, a core member of the Hippo pathway, regulates C2C12 myogenesis. We found that the major components of the mammalian Hippo pathway including Yap are all expressed in skeletal muscles, C2C12 myoblasts and myotubes. In C2C12 myoblasts, Yap Ser127 phosphorylation is low and Yap localises to nuclei. Upon differentiation, Yap Ser127 phosphorylation increases approximately 20-fold and Yap translocates from the nucleus to the cytosol. To test whether the observed increase of Yap Ser127 phosphorylation is required for differentiation we overexpressed hYAP1 S127A, a mutant that can not be phosphorylated at Ser127, in C2C12 myoblasts. We found that overexpression of hYAP S127A prevented myotube formation, whereas the overexpression of wildtype hYAP1 or empty vector had no effect. In addition, more hYAP1 S127A overexpressing cells progressed through the S phase of the cell cycle and the expression of MRFs (myogenin, Myf5), Mef2c and cell cycle regulators (p21, cyclin D1) was significantly changed when compared to wildtype hYAP1 and empty vector overexpressing cells. This data suggests that the phosphorylation of Yap at Ser127 leads to a changed expression of MRFs and cell cycle regulators and is required for C2C12 myoblasts to differentiate into myotubes.
Original languageEnglish
Pages (from-to)619-624
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume393
Issue number4
Early online date10 Feb 2010
DOIs
Publication statusPublished - 19 Mar 2010

Fingerprint

Phosphorylation
Muscle Development
Myoblasts
Skeletal Muscle Fibers
Muscle
Skeletal Muscle
Cell Cycle
Cells
Cyclin-Dependent Kinase Inhibitor p21
Myogenin
Cyclin D1
S Phase
Cytosol

Keywords

  • adaptor proteins, signal transducing
  • animals
  • cell line
  • humans
  • mice
  • mice, inbred C57BL
  • muscle cells
  • muscle development
  • myoblasts
  • myogenic regulatory factors
  • phosphoproteins
  • phosphorylation
  • serine
  • skeletal muscle
  • myogenesis
  • C2C12
  • hippo pathway
  • yap

Cite this

Watt, K. I., Judson, R., Medlow, P., Reid, K., Kurth, T. B., Burniston, J. G., ... Wackerhage, H. (2010). Yap is a novel regulator of C2C12 myogenesis. Biochemical and Biophysical Research Communications, 393(4), 619-624. https://doi.org/10.1016/j.bbrc.2010.02.034

Yap is a novel regulator of C2C12 myogenesis. / Watt, Kevin I; Judson, Robert; Medlow, Paul; Reid, Kenneth; Kurth, Tobias B; Burniston, Jatin G; Ratkevicius, Aivaras; De Bari, Cosimo; Wackerhage, Henning.

In: Biochemical and Biophysical Research Communications, Vol. 393, No. 4, 19.03.2010, p. 619-624.

Research output: Contribution to journalArticle

Watt, KI, Judson, R, Medlow, P, Reid, K, Kurth, TB, Burniston, JG, Ratkevicius, A, De Bari, C & Wackerhage, H 2010, 'Yap is a novel regulator of C2C12 myogenesis', Biochemical and Biophysical Research Communications, vol. 393, no. 4, pp. 619-624. https://doi.org/10.1016/j.bbrc.2010.02.034
Watt KI, Judson R, Medlow P, Reid K, Kurth TB, Burniston JG et al. Yap is a novel regulator of C2C12 myogenesis. Biochemical and Biophysical Research Communications. 2010 Mar 19;393(4):619-624. https://doi.org/10.1016/j.bbrc.2010.02.034
Watt, Kevin I ; Judson, Robert ; Medlow, Paul ; Reid, Kenneth ; Kurth, Tobias B ; Burniston, Jatin G ; Ratkevicius, Aivaras ; De Bari, Cosimo ; Wackerhage, Henning. / Yap is a novel regulator of C2C12 myogenesis. In: Biochemical and Biophysical Research Communications. 2010 ; Vol. 393, No. 4. pp. 619-624.
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abstract = "The expression, regulation and function of mammalian Hippo pathway members in skeletal muscle is largely unknown. The aim of this study was thus to test the hypothesis that core members of the mammalian Hippo pathway are expressed in skeletal muscle and that the transcriptional co-factor Yap, a core member of the Hippo pathway, regulates C2C12 myogenesis. We found that the major components of the mammalian Hippo pathway including Yap are all expressed in skeletal muscles, C2C12 myoblasts and myotubes. In C2C12 myoblasts, Yap Ser127 phosphorylation is low and Yap localises to nuclei. Upon differentiation, Yap Ser127 phosphorylation increases approximately 20-fold and Yap translocates from the nucleus to the cytosol. To test whether the observed increase of Yap Ser127 phosphorylation is required for differentiation we overexpressed hYAP1 S127A, a mutant that can not be phosphorylated at Ser127, in C2C12 myoblasts. We found that overexpression of hYAP S127A prevented myotube formation, whereas the overexpression of wildtype hYAP1 or empty vector had no effect. In addition, more hYAP1 S127A overexpressing cells progressed through the S phase of the cell cycle and the expression of MRFs (myogenin, Myf5), Mef2c and cell cycle regulators (p21, cyclin D1) was significantly changed when compared to wildtype hYAP1 and empty vector overexpressing cells. This data suggests that the phosphorylation of Yap at Ser127 leads to a changed expression of MRFs and cell cycle regulators and is required for C2C12 myoblasts to differentiate into myotubes.",
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AU - Watt, Kevin I

AU - Judson, Robert

AU - Medlow, Paul

AU - Reid, Kenneth

AU - Kurth, Tobias B

AU - Burniston, Jatin G

AU - Ratkevicius, Aivaras

AU - De Bari, Cosimo

AU - Wackerhage, Henning

N1 - Copyright 2010 Elsevier Inc. All rights reserved.

PY - 2010/3/19

Y1 - 2010/3/19

N2 - The expression, regulation and function of mammalian Hippo pathway members in skeletal muscle is largely unknown. The aim of this study was thus to test the hypothesis that core members of the mammalian Hippo pathway are expressed in skeletal muscle and that the transcriptional co-factor Yap, a core member of the Hippo pathway, regulates C2C12 myogenesis. We found that the major components of the mammalian Hippo pathway including Yap are all expressed in skeletal muscles, C2C12 myoblasts and myotubes. In C2C12 myoblasts, Yap Ser127 phosphorylation is low and Yap localises to nuclei. Upon differentiation, Yap Ser127 phosphorylation increases approximately 20-fold and Yap translocates from the nucleus to the cytosol. To test whether the observed increase of Yap Ser127 phosphorylation is required for differentiation we overexpressed hYAP1 S127A, a mutant that can not be phosphorylated at Ser127, in C2C12 myoblasts. We found that overexpression of hYAP S127A prevented myotube formation, whereas the overexpression of wildtype hYAP1 or empty vector had no effect. In addition, more hYAP1 S127A overexpressing cells progressed through the S phase of the cell cycle and the expression of MRFs (myogenin, Myf5), Mef2c and cell cycle regulators (p21, cyclin D1) was significantly changed when compared to wildtype hYAP1 and empty vector overexpressing cells. This data suggests that the phosphorylation of Yap at Ser127 leads to a changed expression of MRFs and cell cycle regulators and is required for C2C12 myoblasts to differentiate into myotubes.

AB - The expression, regulation and function of mammalian Hippo pathway members in skeletal muscle is largely unknown. The aim of this study was thus to test the hypothesis that core members of the mammalian Hippo pathway are expressed in skeletal muscle and that the transcriptional co-factor Yap, a core member of the Hippo pathway, regulates C2C12 myogenesis. We found that the major components of the mammalian Hippo pathway including Yap are all expressed in skeletal muscles, C2C12 myoblasts and myotubes. In C2C12 myoblasts, Yap Ser127 phosphorylation is low and Yap localises to nuclei. Upon differentiation, Yap Ser127 phosphorylation increases approximately 20-fold and Yap translocates from the nucleus to the cytosol. To test whether the observed increase of Yap Ser127 phosphorylation is required for differentiation we overexpressed hYAP1 S127A, a mutant that can not be phosphorylated at Ser127, in C2C12 myoblasts. We found that overexpression of hYAP S127A prevented myotube formation, whereas the overexpression of wildtype hYAP1 or empty vector had no effect. In addition, more hYAP1 S127A overexpressing cells progressed through the S phase of the cell cycle and the expression of MRFs (myogenin, Myf5), Mef2c and cell cycle regulators (p21, cyclin D1) was significantly changed when compared to wildtype hYAP1 and empty vector overexpressing cells. This data suggests that the phosphorylation of Yap at Ser127 leads to a changed expression of MRFs and cell cycle regulators and is required for C2C12 myoblasts to differentiate into myotubes.

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KW - animals

KW - cell line

KW - humans

KW - mice

KW - mice, inbred C57BL

KW - muscle cells

KW - muscle development

KW - myoblasts

KW - myogenic regulatory factors

KW - phosphoproteins

KW - phosphorylation

KW - serine

KW - skeletal muscle

KW - myogenesis

KW - C2C12

KW - hippo pathway

KW - yap

U2 - 10.1016/j.bbrc.2010.02.034

DO - 10.1016/j.bbrc.2010.02.034

M3 - Article

C2 - 20153295

VL - 393

SP - 619

EP - 624

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

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ER -