Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells

Autores
Buitrago, Claudia Graciela; González Pardo, María Verónica; Ronda, Ana Carolina; Boland, Ricardo Leopoldo
Año de publicación
2003
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
We have previously shown that stimulation of proliferation of avian embryonic muscle cells (myoblasts) by 1α,25(OH)2-vitamin D3 (lα,25(OH)2D3) is mediated by activation of the mitogen-activated protein kinase (MAPK; ERK1/2). To understand how lα,25(OH)2D3 up-regulates the MAPK cascade, we have investigated whether the hormone acts upstream through stimulation of Raf-1 and the signaling mechanism by which this effect might take place. Treatment of chick myoblasts with lα,25(OH)2D3 (1 nM) caused a fast increase of Raf-1 serine phosphorylation (1- and 3-fold over basal at 1 and 2 min, respectively), indicating activation of Raf-1 by the hormone. These effects were abolished by preincubation of cells with a specific Ras inhibitor peptide that involves Ras in lα,25(OH)2D3 stimulation of Raf-1. lα,25(OH)2D3 rapidly induced tyrosine de-phosphorylation of Ras-GTPase-activating protein, suggesting that inhibition of Ras-GTP hydrolysis is part of the mechanism by which lα,25(OH)2D3 activates Ras in myoblasts. The protein kinase C (PKC) inhibitors calphostin C, bisindolylmaleimide I, and Ro 318220 blocked lα,25(OH)2D3-induced Raf-1 serine phosphorylation, revealing that hormone stimulation of Raf-1 also involves PKC. In addition, transfection of muscle cells with an antisense oligodeoxynucleotide against PKCα mRNA suppressed serine phosphorylation by lα,25(OH)2D3. The increase in MAPK activity and tyrosine phosphorylation caused by lα,25(OH)2D3 could be abolished by Ras inhibitor peptide, compound PD 98059, which prevents the activation of MEK by Raf-1, or incubation of cell lysates before lα,25(OH)2D3 exposure with an anti-Raf-1 antibody. In conclusion, these results demonstrate for the first time in a lα,25(OH)2D3 target cell that activation of Raf-1 via Ras and PKCα-dependent serine phosphorylation plays a central role in hormone stimulation of the MAPK-signaling pathway leading to muscle cell proliferation.
Fil: Buitrago, Claudia Graciela. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina
Fil: González Pardo, María Verónica. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina
Fil: Ronda, Ana Carolina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina
Fil: Boland, Ricardo Leopoldo. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina
Materia
RAF-1
Protein Kinase C
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/73350
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/73350
network_acronym_str CONICETDig
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network_name_str CONICET Digital (CONICET)
spelling Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle CellsBuitrago, Claudia GracielaGonzález Pardo, María VerónicaRonda, Ana CarolinaBoland, Ricardo LeopoldoRAF-1Protein Kinase Chttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1We have previously shown that stimulation of proliferation of avian embryonic muscle cells (myoblasts) by 1α,25(OH)2-vitamin D3 (lα,25(OH)2D3) is mediated by activation of the mitogen-activated protein kinase (MAPK; ERK1/2). To understand how lα,25(OH)2D3 up-regulates the MAPK cascade, we have investigated whether the hormone acts upstream through stimulation of Raf-1 and the signaling mechanism by which this effect might take place. Treatment of chick myoblasts with lα,25(OH)2D3 (1 nM) caused a fast increase of Raf-1 serine phosphorylation (1- and 3-fold over basal at 1 and 2 min, respectively), indicating activation of Raf-1 by the hormone. These effects were abolished by preincubation of cells with a specific Ras inhibitor peptide that involves Ras in lα,25(OH)2D3 stimulation of Raf-1. lα,25(OH)2D3 rapidly induced tyrosine de-phosphorylation of Ras-GTPase-activating protein, suggesting that inhibition of Ras-GTP hydrolysis is part of the mechanism by which lα,25(OH)2D3 activates Ras in myoblasts. The protein kinase C (PKC) inhibitors calphostin C, bisindolylmaleimide I, and Ro 318220 blocked lα,25(OH)2D3-induced Raf-1 serine phosphorylation, revealing that hormone stimulation of Raf-1 also involves PKC. In addition, transfection of muscle cells with an antisense oligodeoxynucleotide against PKCα mRNA suppressed serine phosphorylation by lα,25(OH)2D3. The increase in MAPK activity and tyrosine phosphorylation caused by lα,25(OH)2D3 could be abolished by Ras inhibitor peptide, compound PD 98059, which prevents the activation of MEK by Raf-1, or incubation of cell lysates before lα,25(OH)2D3 exposure with an anti-Raf-1 antibody. In conclusion, these results demonstrate for the first time in a lα,25(OH)2D3 target cell that activation of Raf-1 via Ras and PKCα-dependent serine phosphorylation plays a central role in hormone stimulation of the MAPK-signaling pathway leading to muscle cell proliferation.Fil: Buitrago, Claudia Graciela. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; ArgentinaFil: González Pardo, María Verónica. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; ArgentinaFil: Ronda, Ana Carolina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; ArgentinaFil: Boland, Ricardo Leopoldo. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; ArgentinaAmerican Society for Biochemistry and Molecular Biology2003-01-24info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/73350Buitrago, Claudia Graciela; González Pardo, María Verónica; Ronda, Ana Carolina; Boland, Ricardo Leopoldo; Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 278; 4; 24-1-2003; 2199-22050021-92581083-351XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/278/4/2199.fullinfo:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M205732200info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:03:57Zoai:ri.conicet.gov.ar:11336/73350instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:03:58.09CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells
title Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells
spellingShingle Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells
Buitrago, Claudia Graciela
RAF-1
Protein Kinase C
title_short Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells
title_full Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells
title_fullStr Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells
title_full_unstemmed Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells
title_sort Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells
dc.creator.none.fl_str_mv Buitrago, Claudia Graciela
González Pardo, María Verónica
Ronda, Ana Carolina
Boland, Ricardo Leopoldo
author Buitrago, Claudia Graciela
author_facet Buitrago, Claudia Graciela
González Pardo, María Verónica
Ronda, Ana Carolina
Boland, Ricardo Leopoldo
author_role author
author2 González Pardo, María Verónica
Ronda, Ana Carolina
Boland, Ricardo Leopoldo
author2_role author
author
author
dc.subject.none.fl_str_mv RAF-1
Protein Kinase C
topic RAF-1
Protein Kinase C
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv We have previously shown that stimulation of proliferation of avian embryonic muscle cells (myoblasts) by 1α,25(OH)2-vitamin D3 (lα,25(OH)2D3) is mediated by activation of the mitogen-activated protein kinase (MAPK; ERK1/2). To understand how lα,25(OH)2D3 up-regulates the MAPK cascade, we have investigated whether the hormone acts upstream through stimulation of Raf-1 and the signaling mechanism by which this effect might take place. Treatment of chick myoblasts with lα,25(OH)2D3 (1 nM) caused a fast increase of Raf-1 serine phosphorylation (1- and 3-fold over basal at 1 and 2 min, respectively), indicating activation of Raf-1 by the hormone. These effects were abolished by preincubation of cells with a specific Ras inhibitor peptide that involves Ras in lα,25(OH)2D3 stimulation of Raf-1. lα,25(OH)2D3 rapidly induced tyrosine de-phosphorylation of Ras-GTPase-activating protein, suggesting that inhibition of Ras-GTP hydrolysis is part of the mechanism by which lα,25(OH)2D3 activates Ras in myoblasts. The protein kinase C (PKC) inhibitors calphostin C, bisindolylmaleimide I, and Ro 318220 blocked lα,25(OH)2D3-induced Raf-1 serine phosphorylation, revealing that hormone stimulation of Raf-1 also involves PKC. In addition, transfection of muscle cells with an antisense oligodeoxynucleotide against PKCα mRNA suppressed serine phosphorylation by lα,25(OH)2D3. The increase in MAPK activity and tyrosine phosphorylation caused by lα,25(OH)2D3 could be abolished by Ras inhibitor peptide, compound PD 98059, which prevents the activation of MEK by Raf-1, or incubation of cell lysates before lα,25(OH)2D3 exposure with an anti-Raf-1 antibody. In conclusion, these results demonstrate for the first time in a lα,25(OH)2D3 target cell that activation of Raf-1 via Ras and PKCα-dependent serine phosphorylation plays a central role in hormone stimulation of the MAPK-signaling pathway leading to muscle cell proliferation.
Fil: Buitrago, Claudia Graciela. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina
Fil: González Pardo, María Verónica. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina
Fil: Ronda, Ana Carolina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina
Fil: Boland, Ricardo Leopoldo. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina
description We have previously shown that stimulation of proliferation of avian embryonic muscle cells (myoblasts) by 1α,25(OH)2-vitamin D3 (lα,25(OH)2D3) is mediated by activation of the mitogen-activated protein kinase (MAPK; ERK1/2). To understand how lα,25(OH)2D3 up-regulates the MAPK cascade, we have investigated whether the hormone acts upstream through stimulation of Raf-1 and the signaling mechanism by which this effect might take place. Treatment of chick myoblasts with lα,25(OH)2D3 (1 nM) caused a fast increase of Raf-1 serine phosphorylation (1- and 3-fold over basal at 1 and 2 min, respectively), indicating activation of Raf-1 by the hormone. These effects were abolished by preincubation of cells with a specific Ras inhibitor peptide that involves Ras in lα,25(OH)2D3 stimulation of Raf-1. lα,25(OH)2D3 rapidly induced tyrosine de-phosphorylation of Ras-GTPase-activating protein, suggesting that inhibition of Ras-GTP hydrolysis is part of the mechanism by which lα,25(OH)2D3 activates Ras in myoblasts. The protein kinase C (PKC) inhibitors calphostin C, bisindolylmaleimide I, and Ro 318220 blocked lα,25(OH)2D3-induced Raf-1 serine phosphorylation, revealing that hormone stimulation of Raf-1 also involves PKC. In addition, transfection of muscle cells with an antisense oligodeoxynucleotide against PKCα mRNA suppressed serine phosphorylation by lα,25(OH)2D3. The increase in MAPK activity and tyrosine phosphorylation caused by lα,25(OH)2D3 could be abolished by Ras inhibitor peptide, compound PD 98059, which prevents the activation of MEK by Raf-1, or incubation of cell lysates before lα,25(OH)2D3 exposure with an anti-Raf-1 antibody. In conclusion, these results demonstrate for the first time in a lα,25(OH)2D3 target cell that activation of Raf-1 via Ras and PKCα-dependent serine phosphorylation plays a central role in hormone stimulation of the MAPK-signaling pathway leading to muscle cell proliferation.
publishDate 2003
dc.date.none.fl_str_mv 2003-01-24
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/73350
Buitrago, Claudia Graciela; González Pardo, María Verónica; Ronda, Ana Carolina; Boland, Ricardo Leopoldo; Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 278; 4; 24-1-2003; 2199-2205
0021-9258
1083-351X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/73350
identifier_str_mv Buitrago, Claudia Graciela; González Pardo, María Verónica; Ronda, Ana Carolina; Boland, Ricardo Leopoldo; Activation of RAF-1 through Ras and Protein Kinase Cα Mediates 1α,25(OH)2-Vitamin D3 Regulation of the Mitogen-activated Protein Kinase Pathway in Muscle Cells; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 278; 4; 24-1-2003; 2199-2205
0021-9258
1083-351X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/278/4/2199.full
info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M205732200
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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