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Abstract:

Protein kinase A (PKA), in yeast, plays a major role in controlling metabolism and gene expression in connection with the available nutrient conditions. We here measure, for the first time, a transient change in the in vivo PKA activity, along a cAMP peak produced by 100 mM glucose addition to glycerol-growing cells as well as a change in the phosphorylation state of its catalytic subunit (Tpk1p) following PKA activation. PKA activity was measured in situ in permeabilized cells, preserving its intracellular localization. Comparison of total PKA activity, measured in situ in permeabilized cells with data obtained from in vitro assays in crude extracts, underscores the inhibitory potency of the regulatory subunit within the cell. Tpk1p phosphorylation was detected through non-denaturing gel electrophoresis. Phosphorylation of Tpk1p increases its specificity constant toward kemptide substrate. The use of mutants of the cAMP pathway showed that phosphorylation depends on the activation of PKA via the G-protein coupled receptor pathway triggered by glucose. The phosphorylation state of Tpk1p was followed during the diauxic shift. Tpk1p phosphorylation is dynamic and reversible: its up-regulation correlates with a fully fermentative metabolism, while its down-regulation with stationary phase or respiratory metabolism. Reversible phosphorylation can thus be considered a new control mechanism possibly pointing to a fine-tuning of PKA activity in response to environmental conditions. © 2005 Elsevier Inc. All rights reserved.

Registro:

Documento: Artículo
Título:Glucose-dependent activation of protein kinase A activity in Saccharomyces cerevisiae and phosphorylation of its TPK1 catalytic subunit
Autor:Portela, P.; Moreno, S.
Filiación:Departamento de Química Biológica, Facultad de Ciencias Exactas Y Naturales, Pabellón 2, Buenos Aires 1428, Argentina
Palabras clave:cAMP signalling; Glucose; In situ assay; Phosphorylation; PKA (protein kinase A); Yeast; cyclic AMP dependent protein kinase; G protein coupled receptor; glucose; glycerol; animal cell; article; breathing; catalysis; cell membrane permeability; cell metabolism; down regulation; extract; gel electrophoresis; in vivo study; nonhuman; priority journal; protein phosphorylation; Saccharomyces cerevisiae; upregulation; Catalytic Domain; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Enzyme Activation; Glucose; Glycerol; Mutation; Oligopeptides; Phosphorylation; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Signal Transduction; Animalia; Saccharomyces cerevisiae
Año:2006
Volumen:18
Número:7
Página de inicio:1072
Página de fin:1086
DOI: http://dx.doi.org/10.1016/j.cellsig.2005.09.001
Título revista:Cellular Signalling
Título revista abreviado:Cell. Signal.
ISSN:08986568
CODEN:CESIE
CAS:glucose, 50-99-7, 84778-64-3; glycerol, 56-81-5; Cyclic AMP, 60-92-4; Cyclic AMP-Dependent Protein Kinases, EC 2.7.1.37; Glucose, 50-99-7; Glycerol, 56-81-5; kemptide, 65189-71-1; Oligopeptides; Saccharomyces cerevisiae Proteins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_08986568_v18_n7_p1072_Portela

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Citas:

---------- APA ----------
Portela, P. & Moreno, S. (2006) . Glucose-dependent activation of protein kinase A activity in Saccharomyces cerevisiae and phosphorylation of its TPK1 catalytic subunit. Cellular Signalling, 18(7), 1072-1086.
http://dx.doi.org/10.1016/j.cellsig.2005.09.001
---------- CHICAGO ----------
Portela, P., Moreno, S. "Glucose-dependent activation of protein kinase A activity in Saccharomyces cerevisiae and phosphorylation of its TPK1 catalytic subunit" . Cellular Signalling 18, no. 7 (2006) : 1072-1086.
http://dx.doi.org/10.1016/j.cellsig.2005.09.001
---------- MLA ----------
Portela, P., Moreno, S. "Glucose-dependent activation of protein kinase A activity in Saccharomyces cerevisiae and phosphorylation of its TPK1 catalytic subunit" . Cellular Signalling, vol. 18, no. 7, 2006, pp. 1072-1086.
http://dx.doi.org/10.1016/j.cellsig.2005.09.001
---------- VANCOUVER ----------
Portela, P., Moreno, S. Glucose-dependent activation of protein kinase A activity in Saccharomyces cerevisiae and phosphorylation of its TPK1 catalytic subunit. Cell. Signal. 2006;18(7):1072-1086.
http://dx.doi.org/10.1016/j.cellsig.2005.09.001