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Functional expression of phosphagen kinase systems confers resistance to transient stresses in Saccharomyces cerevisiae by buffering the ATP pool.

Abstract
Phosphagen kinase systems provide different advantages to tissues with high and fluctuating energy demands, in particular an efficient energy buffering system. In this study we show for the first time functional expression of two phosphagen kinase systems in Saccharomyces cerevisiae, which does not normally contain such systems. First, to establish the creatine kinase system, in addition to overexpressing creatine kinase isoenzymes, we had to install the biosynthesis pathway of creatine by co-overexpression of L-arginine:glycine amidinotransferase and guanidinoacetate methyltransferase. Although we could achieve considerable creatine kinase activity, together with more than 3 mM intracellular creatine, this was not sufficient to confer an obvious advantage to the yeast under the specific stress conditions examined here. Second, using arginine kinase, we successfully installed an intracellular phosphagen pool of about 5 mM phosphoarginine. Such arginine kinase-expressing yeast showed improved resistance under two stress challenges that drain cellular energy, which were transient pH reduction and starvation. Although transient starvation led to 50% reduced intracellular ATP concentrations in wild-type yeast, arginine kinase overexpression stabilized the ATP pool at the pre-stress level. Thus, our results demonstrate that temporal energy buffering is an intrinsic property of phosphagen kinases that can be transferred to phylogenetically very distant organisms.
AuthorsFabrizio Canonaco, Uwe Schlattner, Pamela S Pruett, Theo Wallimann, Uwe Sauer
JournalThe Journal of biological chemistry (J Biol Chem) Vol. 277 Issue 35 Pg. 31303-9 (Aug 30 2002) ISSN: 0021-9258 [Print] United States
PMID12036963 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)
Chemical References
  • Carbon Dioxide
  • Adenosine Triphosphate
  • Arginine
  • Creatine Kinase
  • Arginine Kinase
Topics
  • Adenosine Triphosphate (metabolism)
  • Arginine (metabolism)
  • Arginine Kinase (genetics, metabolism)
  • Carbon Dioxide (metabolism)
  • Creatine Kinase (genetics, metabolism)
  • Hydrogen-Ion Concentration
  • Kinetics
  • Models, Biological
  • Saccharomyces cerevisiae (enzymology, growth & development)

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