Crithidia luciliae: effect of purine starvation on S-adenosyl-L-methionine uptake and protein methylation

Exp Parasitol. 1995 Dec;81(4):519-28. doi: 10.1006/expr.1995.1145.

Abstract

The utilization of S-adenosyl-L-[methyl-3H]methionine ([3H-methyl]AdoMet) by Crithidia luciliae was assessed under nutrient-replete and purine-starvation conditions. Uptake experiments with intact cells demonstrated that the radiolabel from this molecule was accumulated by purine-starved organisms at a rate approximately 10-fold greater than that observed in those cultivated in nutrient-replete medium. Purine-starved cells also incorporated the radiolabel into trichloroacetic acid insoluble material at an approximately 10-fold faster rate than nutrient-replete cells. No differences, however, were observed in the intracellular levels of AdoMet and its metabolites between organisms cultivated under the two conditions. Results of comparative labeling studies with [3H-methyl]AdoMet, S-adenosyl-L-[carboxyl-14C]methionine, L-[methyl-3H]methionine and L-[35S]methionine in the presence and absence of cycloheximide demonstrated that the incorporation of label from [3H-methyl]AdoMet was due to transmethylation and was independent of protein synthesis. Further, approximately 15 methylated protein bands were identified by SDS-PAGE analysis. Lysates from both purine-starved and nutrient-replete organisms demonstrated similar levels of activity of three protein methyltransferases (PMI, II, III). The differences observed in [3H-methyl]AdoMet utilization between purine-starved and nutrient-replete C. luciliae may reflect the enhanced purine transport capacity which results from purine starvation.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine / analogs & derivatives
  • Animals
  • Crithidia / drug effects
  • Crithidia / enzymology
  • Crithidia / metabolism*
  • Cycloheximide / pharmacology
  • Decarboxylation
  • Methionine / metabolism
  • Methylation
  • Protein Methyltransferases / metabolism
  • Protein Processing, Post-Translational*
  • Protein Synthesis Inhibitors / pharmacology
  • Purines / metabolism*
  • S-Adenosylhomocysteine / metabolism
  • S-Adenosylmethionine / analogs & derivatives
  • S-Adenosylmethionine / metabolism*
  • Thionucleosides / metabolism

Substances

  • Protein Synthesis Inhibitors
  • Purines
  • Thionucleosides
  • S-Adenosylmethionine
  • S-Adenosylhomocysteine
  • Cycloheximide
  • Methionine
  • Protein Methyltransferases
  • Adenosine