Regulation of PKA activity by an autophosphorylation mechanism in Saccharomyces cerevisiae

PKA (cAMP-dependent protein kinase) activity, as well as that of other AGC members, is regulated by multiple phosphorylations of its catalytic subunits. In Saccharomyces cerevisiae, the PKA regulatory subunit is encoded by the gene BCY1, and the catalytic subunits are encoded by three genes: TPK1, T...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Tudisca, Vanesa Romina, Nadra, Alejandro Daniel, Moreno, Silvia, Portela, Paula
Publicado:	2014
Materias:	CAMP-dependent protein kinase (PKA) Phosphorylation Saccharomyces cerevisiae Tpk1 cyclic AMP dependent protein kinase glucose holoenzyme article autophosphorylation cell stress cell survival controlled study enzyme activation enzyme active site enzyme phosphorylation enzyme regulation in vitro study in vivo study nonhuman priority journal Catalytic Domain Cyclic AMP-Dependent Protein Kinases Fermentation Glucose Saccharomyces cerevisiae Proteins Serine
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v462_n3_p567_Solari http://hdl.handle.net/20.500.12110/paper_02646021_v462_n3_p567_Solari
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_02646021_v462_n3_p567_Solari
record_format	dspace
spelling	paper:paper_02646021_v462_n3_p567_Solari2023-06-08T15:23:15Z Regulation of PKA activity by an autophosphorylation mechanism in Saccharomyces cerevisiae Tudisca, Vanesa Romina Nadra, Alejandro Daniel Moreno, Silvia Portela, Paula CAMP-dependent protein kinase (PKA) Phosphorylation Saccharomyces cerevisiae Tpk1 cyclic AMP dependent protein kinase glucose holoenzyme article autophosphorylation cell stress cell survival controlled study enzyme activation enzyme active site enzyme phosphorylation enzyme regulation in vitro study in vivo study nonhuman priority journal Saccharomyces cerevisiae Catalytic Domain Cyclic AMP-Dependent Protein Kinases Fermentation Glucose Phosphorylation Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Serine PKA (cAMP-dependent protein kinase) activity, as well as that of other AGC members, is regulated by multiple phosphorylations of its catalytic subunits. In Saccharomyces cerevisiae, the PKA regulatory subunit is encoded by the gene BCY1, and the catalytic subunits are encoded by three genes: TPK1, TPK2 and TPK3. Previously,we have reported that, following cAMP/PKA pathway activation, Tpk1 increases its phosphorylation status. Now, in vivo genetic and in vitro experiments indicate an autophosphorylation mechanism for Tpk1. Using array peptides derived from Tpk1, we identified Ser179 as a target residue. Tpk1 is phosphorylated on Ser179 in vivo during glucose stimulus. Reduction of the activation loop Thr241 phosphorylation increases Ser179 autophosphorylation. To evaluate the role of phosphorylation on Ser 179, wemade strains expressing tpk1S179A or tpk1 S179D as the sole PKA kinase source. Our results suggest that Ser179 phosphorylation increases the reactivity towards the substrate without affecting the formation of the holoenzyme. Phenotypic readout analysis showed that Ser179 phosphorylation increases in vivo PKA activity, reducingcell survival, stress and lifespan. Ser179 phosphorylation increases Tpk1 cytoplasmic accumulation in glucose-grown cells. These results describe for the first time that an autophosphorylation mechanism on Tpk1 controls PKA activity in response to glucose availability. © 2014 Biochemical Society. Fil:Tudisca, V. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Nadra, A.D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Moreno, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Portela, P. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2014 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v462_n3_p567_Solari http://hdl.handle.net/20.500.12110/paper_02646021_v462_n3_p567_Solari
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	CAMP-dependent protein kinase (PKA) Phosphorylation Saccharomyces cerevisiae Tpk1 cyclic AMP dependent protein kinase glucose holoenzyme article autophosphorylation cell stress cell survival controlled study enzyme activation enzyme active site enzyme phosphorylation enzyme regulation in vitro study in vivo study nonhuman priority journal Saccharomyces cerevisiae Catalytic Domain Cyclic AMP-Dependent Protein Kinases Fermentation Glucose Phosphorylation Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Serine
spellingShingle	CAMP-dependent protein kinase (PKA) Phosphorylation Saccharomyces cerevisiae Tpk1 cyclic AMP dependent protein kinase glucose holoenzyme article autophosphorylation cell stress cell survival controlled study enzyme activation enzyme active site enzyme phosphorylation enzyme regulation in vitro study in vivo study nonhuman priority journal Saccharomyces cerevisiae Catalytic Domain Cyclic AMP-Dependent Protein Kinases Fermentation Glucose Phosphorylation Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Serine Tudisca, Vanesa Romina Nadra, Alejandro Daniel Moreno, Silvia Portela, Paula Regulation of PKA activity by an autophosphorylation mechanism in Saccharomyces cerevisiae
topic_facet	CAMP-dependent protein kinase (PKA) Phosphorylation Saccharomyces cerevisiae Tpk1 cyclic AMP dependent protein kinase glucose holoenzyme article autophosphorylation cell stress cell survival controlled study enzyme activation enzyme active site enzyme phosphorylation enzyme regulation in vitro study in vivo study nonhuman priority journal Saccharomyces cerevisiae Catalytic Domain Cyclic AMP-Dependent Protein Kinases Fermentation Glucose Phosphorylation Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Serine
description	PKA (cAMP-dependent protein kinase) activity, as well as that of other AGC members, is regulated by multiple phosphorylations of its catalytic subunits. In Saccharomyces cerevisiae, the PKA regulatory subunit is encoded by the gene BCY1, and the catalytic subunits are encoded by three genes: TPK1, TPK2 and TPK3. Previously,we have reported that, following cAMP/PKA pathway activation, Tpk1 increases its phosphorylation status. Now, in vivo genetic and in vitro experiments indicate an autophosphorylation mechanism for Tpk1. Using array peptides derived from Tpk1, we identified Ser179 as a target residue. Tpk1 is phosphorylated on Ser179 in vivo during glucose stimulus. Reduction of the activation loop Thr241 phosphorylation increases Ser179 autophosphorylation. To evaluate the role of phosphorylation on Ser 179, wemade strains expressing tpk1S179A or tpk1 S179D as the sole PKA kinase source. Our results suggest that Ser179 phosphorylation increases the reactivity towards the substrate without affecting the formation of the holoenzyme. Phenotypic readout analysis showed that Ser179 phosphorylation increases in vivo PKA activity, reducingcell survival, stress and lifespan. Ser179 phosphorylation increases Tpk1 cytoplasmic accumulation in glucose-grown cells. These results describe for the first time that an autophosphorylation mechanism on Tpk1 controls PKA activity in response to glucose availability. © 2014 Biochemical Society.
author	Tudisca, Vanesa Romina Nadra, Alejandro Daniel Moreno, Silvia Portela, Paula
author_facet	Tudisca, Vanesa Romina Nadra, Alejandro Daniel Moreno, Silvia Portela, Paula
author_sort	Tudisca, Vanesa Romina
title	Regulation of PKA activity by an autophosphorylation mechanism in Saccharomyces cerevisiae
title_short	Regulation of PKA activity by an autophosphorylation mechanism in Saccharomyces cerevisiae
title_full	Regulation of PKA activity by an autophosphorylation mechanism in Saccharomyces cerevisiae
title_fullStr	Regulation of PKA activity by an autophosphorylation mechanism in Saccharomyces cerevisiae
title_full_unstemmed	Regulation of PKA activity by an autophosphorylation mechanism in Saccharomyces cerevisiae
title_sort	regulation of pka activity by an autophosphorylation mechanism in saccharomyces cerevisiae
publishDate	2014
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v462_n3_p567_Solari http://hdl.handle.net/20.500.12110/paper_02646021_v462_n3_p567_Solari
work_keys_str_mv	AT tudiscavanesaromina regulationofpkaactivitybyanautophosphorylationmechanisminsaccharomycescerevisiae AT nadraalejandrodaniel regulationofpkaactivitybyanautophosphorylationmechanisminsaccharomycescerevisiae AT morenosilvia regulationofpkaactivitybyanautophosphorylationmechanisminsaccharomycescerevisiae AT portelapaula regulationofpkaactivitybyanautophosphorylationmechanisminsaccharomycescerevisiae
_version_	1768542033832574976

Regulation of PKA activity by an autophosphorylation mechanism in Saccharomyces cerevisiae

Ejemplares similares