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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todo:paper_15671356_v3_n1_p119_Portela2023-10-03T16:26:34Z Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo Portela, P. Van Dijck, P. Thevelein, J.M. Moreno, S. 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. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar 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 |
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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, P. Van Dijck, P. Thevelein, J.M. Moreno, S. 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. |
| format |
JOUR |
| author |
Portela, P. Van Dijck, P. Thevelein, J.M. Moreno, S. |
| author_facet |
Portela, P. Van Dijck, P. Thevelein, J.M. Moreno, S. |
| author_sort |
Portela, P. |
| 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 |
| url |
http://hdl.handle.net/20.500.12110/paper_15671356_v3_n1_p119_Portela |
| work_keys_str_mv |
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| _version_ |
1807316573656973312 |