Artículo

La versión final de este artículo es de uso interno de la institución.
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

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.

Registro:

Documento: Artículo
Título:Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo
Autor:Portela, P.; Van Dijck, P.; Thevelein, J.M.; Moreno, S.
Filiación:Depto. de Quím. Biol., Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires, 1428 Buenos Aires, Argentina
Lab. voor Moleculaire Celbiologie, Vlaams Instituut voor Biotechnologie, Katholieke Universiteit Leuven, Kasteelpark Arenberg 31, 3001 Leuven-Heverlee, Belgium
Palabras clave: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
Año:2003
Volumen:3
Número:1
Página de inicio:119
Página de fin:126
DOI: http://dx.doi.org/10.1016/S1567-1356(02)00158-7
Título revista:FEMS Yeast Research
Título revista abreviado:FEMS Yeast Res.
ISSN:15671356
CODEN:FYREA
CAS:cyclic AMP, 60-92-4; galactose, 26566-61-0, 50855-33-9, 59-23-4; glucose, 50-99-7, 84778-64-3; glycogen, 9005-79-2; protein, 67254-75-5; trehalose, 99-20-7; Cyclic AMP, 60-92-4; Cyclic AMP-Dependent Protein Kinases, EC 2.7.1.37; Galactose, 26566-61-0; Glucose, 50-99-7
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_15671356_v3_n1_p119_Portela

Referencias:

  • Toda, T., Cameron, S., Sass, P., Zoller, M., Scott, J.D., Mc Bullen, B., Hurwitz, M., Wigler, M., Cloning and characterization of BCY1, a locus encoding a regulatory subunit of the cyclic AMP-dependent protein kinase in Saccharomyces cerevisiae (1987) Mol. Cell. Biol., 7, pp. 1371-1377
  • Toda, T., Cameron, S., Sass, P., Zoller, M., Wigler, M., Three different genes in Saccharomyces cerevisiae encode the catalytic subunits of the cAMP-dependent protein kinase (1987) Cell, 50, pp. 277-287
  • Thevelein, J.M., De Winde, J.H., Novel sensing mechanism and target for the cAMP-protein kinase A pathway in the yeast Saccharomyces cerevisiae (1999) Mol. Microbiol., 33, pp. 904-918
  • Kuret, J., Johnson, K.E., Nicolette, C., Zoller, M.J., Mutagenesis of the regulatory subunit of yeast cyclic AMP-dependent protein kinase. Isolation of site-directed mutants with altered binding affinity for catalytic subunit (1988) J. Biol. Chem., 263, pp. 9149-9154
  • Sorol, M.R., Pereyra, E., Mizyrycki, C., Rossi, S., Moreno, S., Protein kinase A activity in permeabilised cells as an approximation to in vivo activity (2001) Exp. Cell Res., 271, pp. 337-343
  • Zaremberg, V., Moreno, S., Analysis of the mechanism of activation of cAMP-dependent protein kinase through the study of mutants of yeast regulatory subunit (1996) Eur. J. Biochem., 237, pp. 136-142
  • Portela, P., Zaremberg, V., Moreno, S., Evaluation of in vivo activation of protein kinase A under non-dissociable conditions through the overexpression of wild-type and mutant regulatory subunits in Saccharomyces cerevisiae (2001) Microbiology, 147, pp. 1149-1159
  • Ito, H., Fukuda, Y., Murata, K., Kimura, A., Transformation of intact yeast cells treated with alkali cations (1983) J. Bacteriol., 153, pp. 163-168
  • Zähringer, H., Thevelein, J.M., Nwaka, S., Induction of neutral trehalase Nth1 by heat and osmotic stress is controlled by STRE elements Msn2/Msn4 transcription factors: Variations of PKA affect during stress and growth (2000) Mol. Microbiol., 35, pp. 397-406
  • Maggesse, M.C., Galvagno, M.A., Cantore, M.L., Passeron, S., In situ measurement of cAMP related enzymes in the dimorphic fungus Mucor rouxii (1982) Cell Biol. Int. Rep., 6, pp. 1101-1108
  • Roskoski R., Jr., Assay of protein kinase (1983) Methods Enzymol., 99, pp. 3-6
  • Colombo, S., Ma, P., Cauwenberg, L., Winderickx, J., Crauwels, M., Teunissen, A., Nauwelaers, D., Thevelein, J.M., Involvement of distinct G-proteins, Gpa2 and Ras, in glucose- and intracellular acidification-induced cAMP signalling in the yeast Saccharomyces cerevisiae (1998) EMBO J., 17, pp. 3326-3341
  • Rose, M.D., Winston, F., Hieter, P., (1990) Methods in Yeast Genetics, , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
  • Bradford, M.M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding (1976) Anal. Biochem., 72, pp. 248-254
  • Görner, W., Durchschlag, E., Martínez-Pastor, M.T., Estruch, F., Ammerer, G., Hamilton, B., Ruis, H., Schüller, C., Nuclear localization of the C2H2 zinc finger protein Msn2p is regulated by stress and protein kinase A activity (1998) Genes Dev., 12, pp. 586-597
  • Zähringer, H., Holzer, H., Nwaka, S., Stability of neutral trehalase during heat stress in Saccharomyces cerevisiae is dependent on the activity of the catalytic subunits of cAMP-dependent protein kinase, TPK1 and TPK2 (1998) Eur. J. Biochem., 255, pp. 544-551
  • Parrou, J.L., Enjalbert, B., Plourde, L., Bauche, A., Gonzalez, B., François, J., Dynamic responses of reserve carbohydrate metabolism under carbon and nitrogen limitations in Saccharomyces cerevisiae (1999) Yeast, 15, pp. 191-203
  • Cameron, S., Levin, L., Zoller, M., Wigler, M., CAMP-independent control of sporulation, glycogen metabolism and heat shock resistance in Saccharomyces cerevisiae (1988) Cell, 53, pp. 555-566
  • Thevelein, J.M., De Winde, J.H., Novel sensing mechanisms and targets for the cAMP-protein kinase A pathway in the yeast Saccharomyces cerevisiae (1999) Mol. Microbiol., 33, pp. 904-918
  • Thevelein, J.M., The RAS-adenylate cyclase pathway and cell cycle control in Saccharomyces cerevisiae (1992) Antonie Van Leeuwenhoek, 62, pp. 109-130
  • Tatchell, K., RAS genes in the budding yeast Saccharomyces cerevisiae (1993) Signal Transduction Prokaryotic and Simple Eukaryotic Systems, pp. 147-188. , Kurjan, J. and Taylor, B.J., Eds. Academic Press, San Diego, CA
  • Thevelein, J.M., Signal transduction in yeast (1994) Yeast, 10, pp. 1753-1790

Citas:

---------- APA ----------
Portela, P., Van Dijck, P., Thevelein, J.M. & Moreno, S. (2003) . Activation state of protein kinase A as measured in permeabilised Saccharomyces cerevisiae cells correlates with PKA-controlled phenotypes in vivo. FEMS Yeast Research, 3(1), 119-126.
http://dx.doi.org/10.1016/S1567-1356(02)00158-7
---------- CHICAGO ----------
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" . FEMS Yeast Research 3, no. 1 (2003) : 119-126.
http://dx.doi.org/10.1016/S1567-1356(02)00158-7
---------- MLA ----------
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" . FEMS Yeast Research, vol. 3, no. 1, 2003, pp. 119-126.
http://dx.doi.org/10.1016/S1567-1356(02)00158-7
---------- VANCOUVER ----------
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. FEMS Yeast Res. 2003;3(1):119-126.
http://dx.doi.org/10.1016/S1567-1356(02)00158-7