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Low activity of the yeast cAMP-dependent protein kinase catalytic subunit Tpk3 is due to the poor expression of the TPK3 gene

Authors

  • María J. MAZÓN,

    Corresponding author
    1. Instituto de Investigaciones Biomédicas del C. S. I. C., Departamento de Bioquímica, Facultad de Medicina, U. A. M., Madrid, Spain
      Correspondence to M. J. Mazón, Instituto de Investigaciones Biomédicas del C. S. I. C., C/Arturo Duperier n°4, E-28029 Madrid, Spain
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  • M. Margarita BEHRENS,

    1. Instituto de Investigaciones Biomédicas del C. S. I. C., Departamento de Bioquímica, Facultad de Medicina, U. A. M., Madrid, Spain
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  • Eulalia MORGADO,

    1. Instituto de Investigaciones Biomédicas del C. S. I. C., Departamento de Bioquímica, Facultad de Medicina, U. A. M., Madrid, Spain
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  • Francisco PORTILLO

    1. Instituto de Investigaciones Biomédicas del C. S. I. C., Departamento de Bioquímica, Facultad de Medicina, U. A. M., Madrid, Spain
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Correspondence to M. J. Mazón, Instituto de Investigaciones Biomédicas del C. S. I. C., C/Arturo Duperier n°4, E-28029 Madrid, Spain

Abstract

Three genes TPK1, TPK2 and TPK3 encode in Saccharomyces cerevisiae distinct catalytic subunits of cAMP-dependent protein kinase (cAPK). We have measured cAPK activity in vitro and, indirectly, in vivo in yeast strains carrying only one of the three TPK genes. The strain containing TPK3 as the only intact TPK gene showed nearly undetectable phosphorylating activity and no TPK3 mRNA could be detected, although the cells grow normally. Overexpression of TPK3 in a high copy vector or under the control of the inducible GAL1 promoter did not by itself result in a corresponding increase in activity but coexpression of BCY1, the gene coding for the regulatory subunit, was necessary in both cases to achieve high levels of phosphorylating activity. Moreover, BCY1 overexpression not only increased Tpk3 catalytic activity but also increased the amount of TPK3 mRNA detected in Northern blots.

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