Different roles of H-ras for regulation of myosin heavy chain promoters in satellite cell-derived muscle cell culture during proliferation and differentiation

Michael E Scholz, Joachim D Meissner, Renate J Scheibe, Patrick K Umeda, Kin-Chow Chang, Gerolf Gros, Hans-Peter Kubis

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The effect of constitutively activated proto-oncogene H-ras (H-rasQ61L) on the regulation of myosin heavy chain (MHC) promoter activities was investigated in rabbit satellite cell-derived muscle cell culture during the proliferation stage and early and later stages of differentiation, respectively. During proliferation, overexpression of H-rasQ61L did not affect basal level of activity of the slow MHCI/beta or the fast MHCIId/x promoter luciferase reporter gene construct in transient transfection assays. By contrast, H-rasQ61L affected both MHC promoter activities during differentiation, and this effect changes from inactivation after 2 days to activation after 4 days of differentiation. The activating effect of H-rasQ61L on both MHC promoters after 4 days of differentiation was significantly reduced by LY-294002, a specific inhibitor of the phosphoinositol-3-kinase (PI3K), a downstream target of Ras. Furthermore, the protein kinase Akt (protein kinase B), a downstream target of PI3k, was activated 4 days after initiation of differentiation in myotubes overexpressing H-rasQ61L. By contrast, inhibition of another Ras downstream pathway, mitogen-activated protein kinase kinase 1/2-extracellular signal-regulated protein kinase 1/2 (MKK1/2-ERK1/2-MAPK), increased activities of both MHC promoters, indicating a suppressive role of this pathway. Moreover, the Ras-PI3K-Akt signaling pathway is involved in the activation of MHCI/beta and IId/x promoters in a later stage of differentiation of muscle cells, presumably by a known inhibiting effect of activated Akt on the MKK1/2-ERK1/2-MAPK pathway. The experiments demonstrate that during differentiation of muscle cells activated H-ras is an important regulator of MHC isoform promoter function with opposite effects during early and later stages.
Original languageEnglish
Pages (from-to)C1012-C1018
Number of pages7
JournalAmerican Journal of Physiology: Cell Physiology
Volume297
Issue number4
DOIs
Publication statusPublished - 22 Jul 2009

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Myosin Heavy Chains
Muscle Cells
Cell Culture Techniques
Protein Kinases
Phosphotransferases
MAP Kinase Kinase 2
MAP Kinase Kinase 1
Proto-Oncogene Proteins c-akt
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Mitogen-Activated Protein Kinase 3
Proto-Oncogenes
MAP Kinase Signaling System
Skeletal Muscle Fibers
Luciferases
Reporter Genes
Transfection
Protein Isoforms
Rabbits

Keywords

  • 1-phosphatidylinositol 3-kinase
  • animals
  • cell differentiation
  • cell proliferation
  • cells, cultured
  • chromones
  • genes, ras
  • MAP kinase signaling system
  • morpholines
  • myosin heavy chains
  • promoter regions, genetic
  • proto-oncogene proteins p21(ras)
  • rabbits
  • satellite cells, skeletal muscle
  • signal transduction

Cite this

Different roles of H-ras for regulation of myosin heavy chain promoters in satellite cell-derived muscle cell culture during proliferation and differentiation. / Scholz, Michael E; Meissner, Joachim D; Scheibe, Renate J; Umeda, Patrick K; Chang, Kin-Chow; Gros, Gerolf; Kubis, Hans-Peter.

In: American Journal of Physiology: Cell Physiology, Vol. 297, No. 4, 22.07.2009, p. C1012-C1018.

Research output: Contribution to journalArticle

Scholz, Michael E ; Meissner, Joachim D ; Scheibe, Renate J ; Umeda, Patrick K ; Chang, Kin-Chow ; Gros, Gerolf ; Kubis, Hans-Peter. / Different roles of H-ras for regulation of myosin heavy chain promoters in satellite cell-derived muscle cell culture during proliferation and differentiation. In: American Journal of Physiology: Cell Physiology. 2009 ; Vol. 297, No. 4. pp. C1012-C1018.
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T1 - Different roles of H-ras for regulation of myosin heavy chain promoters in satellite cell-derived muscle cell culture during proliferation and differentiation

AU - Scholz, Michael E

AU - Meissner, Joachim D

AU - Scheibe, Renate J

AU - Umeda, Patrick K

AU - Chang, Kin-Chow

AU - Gros, Gerolf

AU - Kubis, Hans-Peter

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N2 - The effect of constitutively activated proto-oncogene H-ras (H-rasQ61L) on the regulation of myosin heavy chain (MHC) promoter activities was investigated in rabbit satellite cell-derived muscle cell culture during the proliferation stage and early and later stages of differentiation, respectively. During proliferation, overexpression of H-rasQ61L did not affect basal level of activity of the slow MHCI/beta or the fast MHCIId/x promoter luciferase reporter gene construct in transient transfection assays. By contrast, H-rasQ61L affected both MHC promoter activities during differentiation, and this effect changes from inactivation after 2 days to activation after 4 days of differentiation. The activating effect of H-rasQ61L on both MHC promoters after 4 days of differentiation was significantly reduced by LY-294002, a specific inhibitor of the phosphoinositol-3-kinase (PI3K), a downstream target of Ras. Furthermore, the protein kinase Akt (protein kinase B), a downstream target of PI3k, was activated 4 days after initiation of differentiation in myotubes overexpressing H-rasQ61L. By contrast, inhibition of another Ras downstream pathway, mitogen-activated protein kinase kinase 1/2-extracellular signal-regulated protein kinase 1/2 (MKK1/2-ERK1/2-MAPK), increased activities of both MHC promoters, indicating a suppressive role of this pathway. Moreover, the Ras-PI3K-Akt signaling pathway is involved in the activation of MHCI/beta and IId/x promoters in a later stage of differentiation of muscle cells, presumably by a known inhibiting effect of activated Akt on the MKK1/2-ERK1/2-MAPK pathway. The experiments demonstrate that during differentiation of muscle cells activated H-ras is an important regulator of MHC isoform promoter function with opposite effects during early and later stages.

AB - The effect of constitutively activated proto-oncogene H-ras (H-rasQ61L) on the regulation of myosin heavy chain (MHC) promoter activities was investigated in rabbit satellite cell-derived muscle cell culture during the proliferation stage and early and later stages of differentiation, respectively. During proliferation, overexpression of H-rasQ61L did not affect basal level of activity of the slow MHCI/beta or the fast MHCIId/x promoter luciferase reporter gene construct in transient transfection assays. By contrast, H-rasQ61L affected both MHC promoter activities during differentiation, and this effect changes from inactivation after 2 days to activation after 4 days of differentiation. The activating effect of H-rasQ61L on both MHC promoters after 4 days of differentiation was significantly reduced by LY-294002, a specific inhibitor of the phosphoinositol-3-kinase (PI3K), a downstream target of Ras. Furthermore, the protein kinase Akt (protein kinase B), a downstream target of PI3k, was activated 4 days after initiation of differentiation in myotubes overexpressing H-rasQ61L. By contrast, inhibition of another Ras downstream pathway, mitogen-activated protein kinase kinase 1/2-extracellular signal-regulated protein kinase 1/2 (MKK1/2-ERK1/2-MAPK), increased activities of both MHC promoters, indicating a suppressive role of this pathway. Moreover, the Ras-PI3K-Akt signaling pathway is involved in the activation of MHCI/beta and IId/x promoters in a later stage of differentiation of muscle cells, presumably by a known inhibiting effect of activated Akt on the MKK1/2-ERK1/2-MAPK pathway. The experiments demonstrate that during differentiation of muscle cells activated H-ras is an important regulator of MHC isoform promoter function with opposite effects during early and later stages.

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KW - animals

KW - cell differentiation

KW - cell proliferation

KW - cells, cultured

KW - chromones

KW - genes, ras

KW - MAP kinase signaling system

KW - morpholines

KW - myosin heavy chains

KW - promoter regions, genetic

KW - proto-oncogene proteins p21(ras)

KW - rabbits

KW - satellite cells, skeletal muscle

KW - signal transduction

U2 - 10.1152/ajpcell.00567.2008

DO - 10.1152/ajpcell.00567.2008

M3 - Article

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VL - 297

SP - C1012-C1018

JO - American Journal of Physiology: Cell Physiology

JF - American Journal of Physiology: Cell Physiology

SN - 0363-6143

IS - 4

ER -