Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization

Autores
Chang, Jen Kuan; Ni, Yang; Han, Liang; Sinnett-Smith, James; Jacamo, Rodrigo; Rey, Osvaldo; Young, Steven H.; Rozengurt, Enrique
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Although PKC-mediated phosphorylation of protein kinase D1 (PKD1) has been extensively characterized, little is known about PKD1 regulation by other upstream kinases. Here we report that stimulation of epithelial or fibroblastic cells with G protein-coupled receptor agonists, including angiotensin II or bombesin, induced rapid and persistent PKD1 phosphorylation at Ser203, a highly conserved residue located within the PKD1 N-terminal domain. Exposure to PKD or PKC family inhibitors did not prevent PKD1 phosphorylation at Ser203, indicating that it is not mediated by autophosphorylation. In contrast, several lines of evidence indicated that the phosphorylation of PKD1 at Ser203 is mediated by kinases of the class I PAK subfamily, specifically 1) exposing cells to four structurally unrelated PAK inhibitors (PF-3758309, FRAX486, FRAX597, and IPA-3) that act via different mechanisms abrogated PKD1 phosphorylation at Ser203, 2) siRNA-mediated knockdown of PAK1 and PAK2 in IEC-18 and Swiss 3T3 cells blunted PKD1 phosphorylation at Ser203, 3) phosphorylation of Ser203 markedly increased in vitro when recombinant PKD1 was incubated with either PAK1 or PAK2 in the presence of ATP. PAK inhibitors did not interfere with G protein-coupled receptor activation-induced rapid translocation of PKD1 to the plasma membrane but strikingly prevented the dissociation of PKD1 from the plasma membrane and blunted the phosphorylation of nuclear targets, including class IIa histone deacetylases. We conclude that PAK-mediated phosphorylation of PKD1 at Ser203 triggers its membrane dissociation and subsequent entry into the nucleus, thereby regulating the phosphorylation of PKD1 nuclear targets, including class IIa histone deacetylases.
Fil: Chang, Jen Kuan. University of California at Los Angeles. School of Medicine; Estados Unidos
Fil: Ni, Yang. University of California at Los Angeles. School of Medicine; Estados Unidos. Shandong University; China
Fil: Han, Liang. University of California at Los Angeles. School of Medicine; Estados Unidos. Xi'an Jiaotong University; China
Fil: Sinnett-Smith, James. University of California at Los Angeles. School of Medicine; Estados Unidos
Fil: Jacamo, Rodrigo. University Of Texas Md Anderson Cancer Center;
Fil: Rey, Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina
Fil: Young, Steven H.. University of California at Los Angeles. School of Medicine; Estados Unidos. University of Texas; Estados Unidos
Fil: Rozengurt, Enrique. University of Texas; Estados Unidos. University of California at Los Angeles. School of Medicine; Estados Unidos
Materia
GPCR
PKD
PAK
CANCER
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:11336/82596

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network_name_str CONICET Digital (CONICET)
spelling Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localizationChang, Jen KuanNi, YangHan, LiangSinnett-Smith, JamesJacamo, RodrigoRey, OsvaldoYoung, Steven H.Rozengurt, EnriqueGPCRPKDPAKCANCERhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Although PKC-mediated phosphorylation of protein kinase D1 (PKD1) has been extensively characterized, little is known about PKD1 regulation by other upstream kinases. Here we report that stimulation of epithelial or fibroblastic cells with G protein-coupled receptor agonists, including angiotensin II or bombesin, induced rapid and persistent PKD1 phosphorylation at Ser203, a highly conserved residue located within the PKD1 N-terminal domain. Exposure to PKD or PKC family inhibitors did not prevent PKD1 phosphorylation at Ser203, indicating that it is not mediated by autophosphorylation. In contrast, several lines of evidence indicated that the phosphorylation of PKD1 at Ser203 is mediated by kinases of the class I PAK subfamily, specifically 1) exposing cells to four structurally unrelated PAK inhibitors (PF-3758309, FRAX486, FRAX597, and IPA-3) that act via different mechanisms abrogated PKD1 phosphorylation at Ser203, 2) siRNA-mediated knockdown of PAK1 and PAK2 in IEC-18 and Swiss 3T3 cells blunted PKD1 phosphorylation at Ser203, 3) phosphorylation of Ser203 markedly increased in vitro when recombinant PKD1 was incubated with either PAK1 or PAK2 in the presence of ATP. PAK inhibitors did not interfere with G protein-coupled receptor activation-induced rapid translocation of PKD1 to the plasma membrane but strikingly prevented the dissociation of PKD1 from the plasma membrane and blunted the phosphorylation of nuclear targets, including class IIa histone deacetylases. We conclude that PAK-mediated phosphorylation of PKD1 at Ser203 triggers its membrane dissociation and subsequent entry into the nucleus, thereby regulating the phosphorylation of PKD1 nuclear targets, including class IIa histone deacetylases.Fil: Chang, Jen Kuan. University of California at Los Angeles. School of Medicine; Estados UnidosFil: Ni, Yang. University of California at Los Angeles. School of Medicine; Estados Unidos. Shandong University; ChinaFil: Han, Liang. University of California at Los Angeles. School of Medicine; Estados Unidos. Xi'an Jiaotong University; ChinaFil: Sinnett-Smith, James. University of California at Los Angeles. School of Medicine; Estados UnidosFil: Jacamo, Rodrigo. University Of Texas Md Anderson Cancer Center;Fil: Rey, Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaFil: Young, Steven H.. University of California at Los Angeles. School of Medicine; Estados Unidos. University of Texas; Estados UnidosFil: Rozengurt, Enrique. University of Texas; Estados Unidos. University of California at Los Angeles. School of Medicine; Estados UnidosAmerican Society for Biochemistry and Molecular Biology2017-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/82596Chang, Jen Kuan; Ni, Yang; Han, Liang; Sinnett-Smith, James; Jacamo, Rodrigo; et al.; Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 292; 23; 6-2017; 9523-95390021-9258CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/cgi/pmidlookup?view=long&pmid=28408623info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M116.771394info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5465480/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:02:26Zoai:ri.conicet.gov.ar:11336/82596instacron: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 10:02:26.407CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization
title Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization
spellingShingle Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization
Chang, Jen Kuan
GPCR
PKD
PAK
CANCER
title_short Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization
title_full Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization
title_fullStr Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization
title_full_unstemmed Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization
title_sort Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization
dc.creator.none.fl_str_mv Chang, Jen Kuan
Ni, Yang
Han, Liang
Sinnett-Smith, James
Jacamo, Rodrigo
Rey, Osvaldo
Young, Steven H.
Rozengurt, Enrique
author Chang, Jen Kuan
author_facet Chang, Jen Kuan
Ni, Yang
Han, Liang
Sinnett-Smith, James
Jacamo, Rodrigo
Rey, Osvaldo
Young, Steven H.
Rozengurt, Enrique
author_role author
author2 Ni, Yang
Han, Liang
Sinnett-Smith, James
Jacamo, Rodrigo
Rey, Osvaldo
Young, Steven H.
Rozengurt, Enrique
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv GPCR
PKD
PAK
CANCER
topic GPCR
PKD
PAK
CANCER
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Although PKC-mediated phosphorylation of protein kinase D1 (PKD1) has been extensively characterized, little is known about PKD1 regulation by other upstream kinases. Here we report that stimulation of epithelial or fibroblastic cells with G protein-coupled receptor agonists, including angiotensin II or bombesin, induced rapid and persistent PKD1 phosphorylation at Ser203, a highly conserved residue located within the PKD1 N-terminal domain. Exposure to PKD or PKC family inhibitors did not prevent PKD1 phosphorylation at Ser203, indicating that it is not mediated by autophosphorylation. In contrast, several lines of evidence indicated that the phosphorylation of PKD1 at Ser203 is mediated by kinases of the class I PAK subfamily, specifically 1) exposing cells to four structurally unrelated PAK inhibitors (PF-3758309, FRAX486, FRAX597, and IPA-3) that act via different mechanisms abrogated PKD1 phosphorylation at Ser203, 2) siRNA-mediated knockdown of PAK1 and PAK2 in IEC-18 and Swiss 3T3 cells blunted PKD1 phosphorylation at Ser203, 3) phosphorylation of Ser203 markedly increased in vitro when recombinant PKD1 was incubated with either PAK1 or PAK2 in the presence of ATP. PAK inhibitors did not interfere with G protein-coupled receptor activation-induced rapid translocation of PKD1 to the plasma membrane but strikingly prevented the dissociation of PKD1 from the plasma membrane and blunted the phosphorylation of nuclear targets, including class IIa histone deacetylases. We conclude that PAK-mediated phosphorylation of PKD1 at Ser203 triggers its membrane dissociation and subsequent entry into the nucleus, thereby regulating the phosphorylation of PKD1 nuclear targets, including class IIa histone deacetylases.
Fil: Chang, Jen Kuan. University of California at Los Angeles. School of Medicine; Estados Unidos
Fil: Ni, Yang. University of California at Los Angeles. School of Medicine; Estados Unidos. Shandong University; China
Fil: Han, Liang. University of California at Los Angeles. School of Medicine; Estados Unidos. Xi'an Jiaotong University; China
Fil: Sinnett-Smith, James. University of California at Los Angeles. School of Medicine; Estados Unidos
Fil: Jacamo, Rodrigo. University Of Texas Md Anderson Cancer Center;
Fil: Rey, Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina
Fil: Young, Steven H.. University of California at Los Angeles. School of Medicine; Estados Unidos. University of Texas; Estados Unidos
Fil: Rozengurt, Enrique. University of Texas; Estados Unidos. University of California at Los Angeles. School of Medicine; Estados Unidos
description Although PKC-mediated phosphorylation of protein kinase D1 (PKD1) has been extensively characterized, little is known about PKD1 regulation by other upstream kinases. Here we report that stimulation of epithelial or fibroblastic cells with G protein-coupled receptor agonists, including angiotensin II or bombesin, induced rapid and persistent PKD1 phosphorylation at Ser203, a highly conserved residue located within the PKD1 N-terminal domain. Exposure to PKD or PKC family inhibitors did not prevent PKD1 phosphorylation at Ser203, indicating that it is not mediated by autophosphorylation. In contrast, several lines of evidence indicated that the phosphorylation of PKD1 at Ser203 is mediated by kinases of the class I PAK subfamily, specifically 1) exposing cells to four structurally unrelated PAK inhibitors (PF-3758309, FRAX486, FRAX597, and IPA-3) that act via different mechanisms abrogated PKD1 phosphorylation at Ser203, 2) siRNA-mediated knockdown of PAK1 and PAK2 in IEC-18 and Swiss 3T3 cells blunted PKD1 phosphorylation at Ser203, 3) phosphorylation of Ser203 markedly increased in vitro when recombinant PKD1 was incubated with either PAK1 or PAK2 in the presence of ATP. PAK inhibitors did not interfere with G protein-coupled receptor activation-induced rapid translocation of PKD1 to the plasma membrane but strikingly prevented the dissociation of PKD1 from the plasma membrane and blunted the phosphorylation of nuclear targets, including class IIa histone deacetylases. We conclude that PAK-mediated phosphorylation of PKD1 at Ser203 triggers its membrane dissociation and subsequent entry into the nucleus, thereby regulating the phosphorylation of PKD1 nuclear targets, including class IIa histone deacetylases.
publishDate 2017
dc.date.none.fl_str_mv 2017-06
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/11336/82596
Chang, Jen Kuan; Ni, Yang; Han, Liang; Sinnett-Smith, James; Jacamo, Rodrigo; et al.; Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 292; 23; 6-2017; 9523-9539
0021-9258
CONICET Digital
CONICET
url http://hdl.handle.net/11336/82596
identifier_str_mv Chang, Jen Kuan; Ni, Yang; Han, Liang; Sinnett-Smith, James; Jacamo, Rodrigo; et al.; Protein kinase D1 (PKD1) phosphorylation on Ser203 by type i p21-activated kinase (PAK) regulates PKD1 localization; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 292; 23; 6-2017; 9523-9539
0021-9258
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/cgi/pmidlookup?view=long&pmid=28408623
info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M116.771394
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5465480/
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/
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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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