Open AccessActivation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA‐controlled phenotypes in vivo
Author(s) -
Portela Paula,
Dijck Patrick,
Thevelein Johan M,
Moreno Silvia
Publication year - 2003
Publication title -
fems yeast research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 92
eISSN - 1567-1364
pISSN - 1567-1356
DOI - 10.1111/j.1567-1364.2003.tb00147.x
Subject(s) - saccharomyces cerevisiae , biology , protein kinase a , in vivo , phenotype , mutant , mutation , biochemistry , galactose , microbiology and biotechnology , kinase , gene , genetics
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.