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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Detalles Bibliográficos
Autores principales: Portela, P., Van Dijck, P., Thevelein, J.M., Moreno, S.
Formato: JOUR
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: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
Descripción
Sumario: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.

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