The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae

Autores
Haesendonckx, Steven; Tudisca, Vanesa Romina; Voordeckers, Karin; Moreno, Silvia Margarita; Thevelein, Johan M.; Portela, Paula
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
PDK1 (phosphoinositide-dependent protein kinase 1) phosphorylates and activates PKA (cAMP-dependent protein kinase) in vitro. Docking of the HM (hydrophobic motif) in the C-terminal tail of the PKA catalytic subunits on to the PIF (PDK1-interacting fragment) pocket of PDK1 is a critical step in this activation process. However, PDK1 regulation of PKA in vivo remains controversial. Saccharomyces cerevisiae contains three PKA catalytic subunits, TPK1, TPK2 and TPK3. We demonstrate that Pkh [PKB (protein kinase B)-activating kinase homologue] protein kinases phosphorylate the activation loop of each Tpk in vivo with various efficiencies. Pkh inactivation reduces the interaction of each catalytic subunit with the regulatory subunit Bcy1 without affecting the specific kinase activity of PKA. Comparative analysis of the in vitro interaction and phosphorylation of Tpks by Pkh1 shows that Tpk1 and Tpk2 interact with Pkh1 through an HM-PIF pocket interaction. Unlike Tpk1, mutagenesis of the activation loop site in Tpk2 does not abolish in vitro phosphorylation, suggesting that Tpk2 contains other, as yet uncharacterized, Pkh1 target sites. Tpk3 is poorly phosphorylated on its activation loop site, and this is due to the weak interaction of Tpk3 with Pkh1 because of the atypical HM found in Tpk3. In conclusion, the results of the present study show that Pkh protein kinases contribute to the divergent regulation of the Tpk catalytic subunits. © The Authors Journal compilation © 2012 Biochemical Society.
Fil: Haesendonckx, Steven. Universidad de Ginebra; Suiza. Katholikie Universiteit Leuven; Bélgica. Flanders Interuniversity Institute For Biotechnology; Bélgica
Fil: Tudisca, Vanesa Romina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Fil: Voordeckers, Karin. Flanders Interuniversity Institute For Biotechnology; Bélgica. Katholikie Universiteit Leuven; Bélgica
Fil: Moreno, Silvia Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Fil: Thevelein, Johan M.. Flanders Interuniversity Institute For Biotechnology; Bélgica. Katholikie Universiteit Leuven; Bélgica
Fil: Portela, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Materia
Bcy1
Camp-Dependent Protein Kinase (Pka)
Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
Pkb (Protein Kinase B)-Activating Kinase Homologue (Pkh)
Saccharomyces Cerevisiae
Tpk
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:11336/66822
Acceder
id CONICETDig_3bf118765652980ce0949688a1fc2173
oai_identifier_str oai:ri.conicet.gov.ar:11336/66822
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiaeHaesendonckx, StevenTudisca, Vanesa RominaVoordeckers, KarinMoreno, Silvia MargaritaThevelein, Johan M.Portela, PaulaBcy1Camp-Dependent Protein Kinase (Pka)Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)Pkb (Protein Kinase B)-Activating Kinase Homologue (Pkh)Saccharomyces CerevisiaeTpkhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1PDK1 (phosphoinositide-dependent protein kinase 1) phosphorylates and activates PKA (cAMP-dependent protein kinase) in vitro. Docking of the HM (hydrophobic motif) in the C-terminal tail of the PKA catalytic subunits on to the PIF (PDK1-interacting fragment) pocket of PDK1 is a critical step in this activation process. However, PDK1 regulation of PKA in vivo remains controversial. Saccharomyces cerevisiae contains three PKA catalytic subunits, TPK1, TPK2 and TPK3. We demonstrate that Pkh [PKB (protein kinase B)-activating kinase homologue] protein kinases phosphorylate the activation loop of each Tpk in vivo with various efficiencies. Pkh inactivation reduces the interaction of each catalytic subunit with the regulatory subunit Bcy1 without affecting the specific kinase activity of PKA. Comparative analysis of the in vitro interaction and phosphorylation of Tpks by Pkh1 shows that Tpk1 and Tpk2 interact with Pkh1 through an HM-PIF pocket interaction. Unlike Tpk1, mutagenesis of the activation loop site in Tpk2 does not abolish in vitro phosphorylation, suggesting that Tpk2 contains other, as yet uncharacterized, Pkh1 target sites. Tpk3 is poorly phosphorylated on its activation loop site, and this is due to the weak interaction of Tpk3 with Pkh1 because of the atypical HM found in Tpk3. In conclusion, the results of the present study show that Pkh protein kinases contribute to the divergent regulation of the Tpk catalytic subunits. © The Authors Journal compilation © 2012 Biochemical Society.Fil: Haesendonckx, Steven. Universidad de Ginebra; Suiza. Katholikie Universiteit Leuven; Bélgica. Flanders Interuniversity Institute For Biotechnology; BélgicaFil: Tudisca, Vanesa Romina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; ArgentinaFil: Voordeckers, Karin. Flanders Interuniversity Institute For Biotechnology; Bélgica. Katholikie Universiteit Leuven; BélgicaFil: Moreno, Silvia Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; ArgentinaFil: Thevelein, Johan M.. Flanders Interuniversity Institute For Biotechnology; Bélgica. Katholikie Universiteit Leuven; BélgicaFil: Portela, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; ArgentinaPortland Press2012-09info: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/11336/66822Haesendonckx, Steven; Tudisca, Vanesa Romina; Voordeckers, Karin; Moreno, Silvia Margarita; Thevelein, Johan M.; et al.; The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae; Portland Press; Biochemical Journal; 448; 3; 9-2012; 307-3200264-6021CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1042/BJ20121061info:eu-repo/semantics/altIdentifier/url/http://www.biochemj.org/content/448/3/307info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:03:20Zoai:ri.conicet.gov.ar:11336/66822instacron: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 10:03:20.967CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae
title The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae
spellingShingle The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae
Haesendonckx, Steven
Bcy1
Camp-Dependent Protein Kinase (Pka)
Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
Pkb (Protein Kinase B)-Activating Kinase Homologue (Pkh)
Saccharomyces Cerevisiae
Tpk
title_short The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae
title_full The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae
title_fullStr The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae
title_full_unstemmed The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae
title_sort The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae
dc.creator.none.fl_str_mv Haesendonckx, Steven
Tudisca, Vanesa Romina
Voordeckers, Karin
Moreno, Silvia Margarita
Thevelein, Johan M.
Portela, Paula
author Haesendonckx, Steven
author_facet Haesendonckx, Steven
Tudisca, Vanesa Romina
Voordeckers, Karin
Moreno, Silvia Margarita
Thevelein, Johan M.
Portela, Paula
author_role author
author2 Tudisca, Vanesa Romina
Voordeckers, Karin
Moreno, Silvia Margarita
Thevelein, Johan M.
Portela, Paula
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Bcy1
Camp-Dependent Protein Kinase (Pka)
Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
Pkb (Protein Kinase B)-Activating Kinase Homologue (Pkh)
Saccharomyces Cerevisiae
Tpk
topic Bcy1
Camp-Dependent Protein Kinase (Pka)
Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
Pkb (Protein Kinase B)-Activating Kinase Homologue (Pkh)
Saccharomyces Cerevisiae
Tpk
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv PDK1 (phosphoinositide-dependent protein kinase 1) phosphorylates and activates PKA (cAMP-dependent protein kinase) in vitro. Docking of the HM (hydrophobic motif) in the C-terminal tail of the PKA catalytic subunits on to the PIF (PDK1-interacting fragment) pocket of PDK1 is a critical step in this activation process. However, PDK1 regulation of PKA in vivo remains controversial. Saccharomyces cerevisiae contains three PKA catalytic subunits, TPK1, TPK2 and TPK3. We demonstrate that Pkh [PKB (protein kinase B)-activating kinase homologue] protein kinases phosphorylate the activation loop of each Tpk in vivo with various efficiencies. Pkh inactivation reduces the interaction of each catalytic subunit with the regulatory subunit Bcy1 without affecting the specific kinase activity of PKA. Comparative analysis of the in vitro interaction and phosphorylation of Tpks by Pkh1 shows that Tpk1 and Tpk2 interact with Pkh1 through an HM-PIF pocket interaction. Unlike Tpk1, mutagenesis of the activation loop site in Tpk2 does not abolish in vitro phosphorylation, suggesting that Tpk2 contains other, as yet uncharacterized, Pkh1 target sites. Tpk3 is poorly phosphorylated on its activation loop site, and this is due to the weak interaction of Tpk3 with Pkh1 because of the atypical HM found in Tpk3. In conclusion, the results of the present study show that Pkh protein kinases contribute to the divergent regulation of the Tpk catalytic subunits. © The Authors Journal compilation © 2012 Biochemical Society.
Fil: Haesendonckx, Steven. Universidad de Ginebra; Suiza. Katholikie Universiteit Leuven; Bélgica. Flanders Interuniversity Institute For Biotechnology; Bélgica
Fil: Tudisca, Vanesa Romina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Fil: Voordeckers, Karin. Flanders Interuniversity Institute For Biotechnology; Bélgica. Katholikie Universiteit Leuven; Bélgica
Fil: Moreno, Silvia Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Fil: Thevelein, Johan M.. Flanders Interuniversity Institute For Biotechnology; Bélgica. Katholikie Universiteit Leuven; Bélgica
Fil: Portela, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
description PDK1 (phosphoinositide-dependent protein kinase 1) phosphorylates and activates PKA (cAMP-dependent protein kinase) in vitro. Docking of the HM (hydrophobic motif) in the C-terminal tail of the PKA catalytic subunits on to the PIF (PDK1-interacting fragment) pocket of PDK1 is a critical step in this activation process. However, PDK1 regulation of PKA in vivo remains controversial. Saccharomyces cerevisiae contains three PKA catalytic subunits, TPK1, TPK2 and TPK3. We demonstrate that Pkh [PKB (protein kinase B)-activating kinase homologue] protein kinases phosphorylate the activation loop of each Tpk in vivo with various efficiencies. Pkh inactivation reduces the interaction of each catalytic subunit with the regulatory subunit Bcy1 without affecting the specific kinase activity of PKA. Comparative analysis of the in vitro interaction and phosphorylation of Tpks by Pkh1 shows that Tpk1 and Tpk2 interact with Pkh1 through an HM-PIF pocket interaction. Unlike Tpk1, mutagenesis of the activation loop site in Tpk2 does not abolish in vitro phosphorylation, suggesting that Tpk2 contains other, as yet uncharacterized, Pkh1 target sites. Tpk3 is poorly phosphorylated on its activation loop site, and this is due to the weak interaction of Tpk3 with Pkh1 because of the atypical HM found in Tpk3. In conclusion, the results of the present study show that Pkh protein kinases contribute to the divergent regulation of the Tpk catalytic subunits. © The Authors Journal compilation © 2012 Biochemical Society.
publishDate 2012
dc.date.none.fl_str_mv 2012-09
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/11336/66822
Haesendonckx, Steven; Tudisca, Vanesa Romina; Voordeckers, Karin; Moreno, Silvia Margarita; Thevelein, Johan M.; et al.; The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae; Portland Press; Biochemical Journal; 448; 3; 9-2012; 307-320
0264-6021
CONICET Digital
CONICET
url http://hdl.handle.net/11336/66822
identifier_str_mv Haesendonckx, Steven; Tudisca, Vanesa Romina; Voordeckers, Karin; Moreno, Silvia Margarita; Thevelein, Johan M.; et al.; The activation loop of PKA catalytic isoforms is differentially phosphorylated by Pkh protein kinases in Saccharomyces cerevisiae; Portland Press; Biochemical Journal; 448; 3; 9-2012; 307-320
0264-6021
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1042/BJ20121061
info:eu-repo/semantics/altIdentifier/url/http://www.biochemj.org/content/448/3/307
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
application/pdf
dc.publisher.none.fl_str_mv Portland Press
publisher.none.fl_str_mv Portland Press
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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