Paradoxical effects of protein synthesis inhibitors on uridine uptake in cultured cells: possible role of uncharged tRNA in regulating metabolism.
Previous studies (J. Biol. Chem, 253: 99--105, 1978) showed that thyrotropin-releasing hormone (TRH) acutely stimulated uridine uptake in pituitary cell (GH 4Cl)) cultures. Studies on the role of protein synthesis in this response to TRH led to the finding that an inhibitor of ribosomal translation, cycloheximide, also stimulated uridine uptake acutely. Studies reported here attempt to determine the mechanism of cycloheximide action and whether cycloheximide and hormone stimulation of uridine uptake occurred by similar pathways. The experiments presented indicate that: (1) seven inhibitors of ribosomal translation stimulated uridine uptake; (2) in contrast, inhibition of protein synthesis at tRNA aminoacylation resulted in reduced rates of uridine uptake; (3) inhibition of tRNA aminoacylation blocked cycloheximide but not TRH stimulation of uptake; (4) cycloheximide stimulation of uptake was restricted to amino acid-depleted cultures; (5) amino acid supplementation stimulated uridine uptake with a time-course identical to that of cycloheximide; (6) cycloheximide and amino acid supplementation promoted reacylation of cellular tRNAs in amino acid-depleted cultures; and (7) cycloheximide stimulation of uridine uptake resulted from enhanced nucleoside phosphorylation rather than increased uridine transport. We conclude that cycloheximide and amino acid stimulation of uridine phosphorylation may be mediated through a common pathway involving the extent of amino-acylation of cellular tRNAs. Furthermore, cycloheximide and TRH stimulate uridine phosphorylation by pathways that are distinguishable. It is apparent that not all cellular effects of cycloheximide can be attributed solely to inhibition of the synthesis of proteins.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology*; D006641:Histidinol / Q000494:pharmacology; D010766:Phosphorylation; D010911:Pituitary Neoplasms; D014176:Protein Biosynthesis; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*; D051381:Rats; D014529:Uridine / Q000378:metabolism*
D000596:Amino Acids; D012346:RNA, Transfer, Amino Acyl; D006641:Histidinol; D000255:Adenosine Triphosphate; D003513:Cycloheximide; D014529:Uridine