Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo

Protein kinase A (PKA) activity was measured in situ in permeabilised Saccharomyces cerevisiae cells in the absence and the presence of cAMP. Four strains genetically predicted to have differential PKA-dependent phenotypes were used: a wild-type strain and a strain containing a bcy1-14 mutation (wit...

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Autores principales:	Portela, Paula, Moreno de Colonna, Silvia
Publicado:	2003
Materias:	cAMP Permeabilisation assay PKA-dependent phenotype Protein kinase A Yeast cyclic AMP cyclic AMP dependent protein kinase galactose glucose glycogen protein protein BCY1 trehalose unclassified drug article cell growth cell permeabilization controlled study correlation analysis enzyme activation enzyme activity enzyme assay fungal cell fungal strain gene activity gene expression gene mutation genotype in vivo study nonhuman phenotype promoter region reporter gene Saccharomyces cerevisiae Cyclic AMP Cyclic AMP-Dependent Protein Kinases Enzyme Activation Galactose Genes, Reporter Glucose Mutation Phenotype Substrate Specificity
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_15671356_v3_n1_p119_Portela http://hdl.handle.net/20.500.12110/paper_15671356_v3_n1_p119_Portela
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_15671356_v3_n1_p119_Portela
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spelling	paper:paper_15671356_v3_n1_p119_Portela2023-06-08T16:24:02Z Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo Portela, Paula Moreno de Colonna, Silvia cAMP Permeabilisation assay PKA-dependent phenotype Protein kinase A Yeast cyclic AMP cyclic AMP dependent protein kinase galactose glucose glycogen protein protein BCY1 trehalose unclassified drug article cell growth cell permeabilization controlled study correlation analysis enzyme activation enzyme activity enzyme assay fungal cell fungal strain gene activity gene expression gene mutation genotype in vivo study nonhuman phenotype promoter region reporter gene Saccharomyces cerevisiae Cyclic AMP Cyclic AMP-Dependent Protein Kinases Enzyme Activation Galactose Genes, Reporter Glucose Mutation Phenotype Saccharomyces cerevisiae Substrate Specificity Saccharomyces cerevisiae Protein kinase A (PKA) activity was measured in situ in permeabilised Saccharomyces cerevisiae cells in the absence and the presence of cAMP. Four strains genetically predicted to have differential PKA-dependent phenotypes were used: a wild-type strain and a strain containing a bcy1-14 mutation (with almost constitutively active PKA), and the same strains with overexpression of the wild-type or mutant BCY1 gene, respectively. Cells were grown on galactose or glucose. The measured phenotypic characteristics were: trehalose and glycogen levels and the activity of a reporter gene under control of the NTH1 promoter. The 'endogenous' PKA activity (measured in situ in the absence of cAMP) showed the best correlation with the PKA-dependent phenotypes determined in vivo. We propose that this parameter offers a good estimate for the degree of activation of PKA in vivo. © 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved. Fil:Portela, P. 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. 2003 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_15671356_v3_n1_p119_Portela http://hdl.handle.net/20.500.12110/paper_15671356_v3_n1_p119_Portela
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 Permeabilisation assay PKA-dependent phenotype Protein kinase A Yeast cyclic AMP cyclic AMP dependent protein kinase galactose glucose glycogen protein protein BCY1 trehalose unclassified drug article cell growth cell permeabilization controlled study correlation analysis enzyme activation enzyme activity enzyme assay fungal cell fungal strain gene activity gene expression gene mutation genotype in vivo study nonhuman phenotype promoter region reporter gene Saccharomyces cerevisiae Cyclic AMP Cyclic AMP-Dependent Protein Kinases Enzyme Activation Galactose Genes, Reporter Glucose Mutation Phenotype Saccharomyces cerevisiae Substrate Specificity Saccharomyces cerevisiae
spellingShingle	cAMP Permeabilisation assay PKA-dependent phenotype Protein kinase A Yeast cyclic AMP cyclic AMP dependent protein kinase galactose glucose glycogen protein protein BCY1 trehalose unclassified drug article cell growth cell permeabilization controlled study correlation analysis enzyme activation enzyme activity enzyme assay fungal cell fungal strain gene activity gene expression gene mutation genotype in vivo study nonhuman phenotype promoter region reporter gene Saccharomyces cerevisiae Cyclic AMP Cyclic AMP-Dependent Protein Kinases Enzyme Activation Galactose Genes, Reporter Glucose Mutation Phenotype Saccharomyces cerevisiae Substrate Specificity Saccharomyces cerevisiae Portela, Paula Moreno de Colonna, Silvia Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo
topic_facet	cAMP Permeabilisation assay PKA-dependent phenotype Protein kinase A Yeast cyclic AMP cyclic AMP dependent protein kinase galactose glucose glycogen protein protein BCY1 trehalose unclassified drug article cell growth cell permeabilization controlled study correlation analysis enzyme activation enzyme activity enzyme assay fungal cell fungal strain gene activity gene expression gene mutation genotype in vivo study nonhuman phenotype promoter region reporter gene Saccharomyces cerevisiae Cyclic AMP Cyclic AMP-Dependent Protein Kinases Enzyme Activation Galactose Genes, Reporter Glucose Mutation Phenotype Saccharomyces cerevisiae Substrate Specificity Saccharomyces cerevisiae
description	Protein kinase A (PKA) activity was measured in situ in permeabilised Saccharomyces cerevisiae cells in the absence and the presence of cAMP. Four strains genetically predicted to have differential PKA-dependent phenotypes were used: a wild-type strain and a strain containing a bcy1-14 mutation (with almost constitutively active PKA), and the same strains with overexpression of the wild-type or mutant BCY1 gene, respectively. Cells were grown on galactose or glucose. The measured phenotypic characteristics were: trehalose and glycogen levels and the activity of a reporter gene under control of the NTH1 promoter. The 'endogenous' PKA activity (measured in situ in the absence of cAMP) showed the best correlation with the PKA-dependent phenotypes determined in vivo. We propose that this parameter offers a good estimate for the degree of activation of PKA in vivo. © 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
author	Portela, Paula Moreno de Colonna, Silvia
author_facet	Portela, Paula Moreno de Colonna, Silvia
author_sort	Portela, Paula
title	Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo
title_short	Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo
title_full	Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo
title_fullStr	Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo
title_full_unstemmed	Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo
title_sort	activation state of protein kinase a as measured in permeabilised saccharomyces cerevisiae cells correlates with pka-controlled phenotypes in vivo
publishDate	2003
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_15671356_v3_n1_p119_Portela http://hdl.handle.net/20.500.12110/paper_15671356_v3_n1_p119_Portela
work_keys_str_mv	AT portelapaula activationstateofproteinkinaseaasmeasuredinpermeabilisedsaccharomycescerevisiaecellscorrelateswithpkacontrolledphenotypesinvivo AT morenodecolonnasilvia activationstateofproteinkinaseaasmeasuredinpermeabilisedsaccharomycescerevisiaecellscorrelateswithpkacontrolledphenotypesinvivo
_version_	1768543390783242240

Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo

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