Estradiol regulation of reactions involved in turnover of the amino acid acceptor terminus of tRNA in the rat uterus

Abstract

Estradiol (E2) increases the specific amino acid acceptor activity of rat uterine tRNAs by increasing the proportion of certain tRNAs with intact and functional 3'-CCA acceptor termini. Activities of tRNA nucleotidyltransferase and 3'-exoribonuclease which synthesize and degrade this terminus, respectively, were measured and neither enzyme was modified by hormone treatment. Since cytidine triphosphate (CTP) levels are below reported Km values for nucleotidyltransferase, changes in CTP concentrations may regulate nucleotidyltransferase activity. An E2-induced 3-fold increase was seen in CTP synthetase activity (conversion of uridine triphosphate, UTP, into CTP). Uterine CTP levels in controls are minute (9 nmol/uterus, approx. 90 microM), and are increased 2.5-fold in E2(12 h)-treated rats. The rate of incorporation of [3H]UTP into the 3'-CCA terminus of tRNA was measured as coupled CTP synthetase-nucleotidyltransferase reactions and a 2.5-fold increase in incorporation occurred 8-12 h after E2 treatment. Injection of azaserine, (inhibitor of CTP synthetase) reduced E2-induced increases in CTP levels, CTP synthetase activity, and leucine acceptor activity of tRNAs. These results indicate that E2 regulates CTP levels by modulation of CTP synthetase activity, and that regulation of synthesis and/or repair of the 3'-CCA terminus of tRNA is proportional to E2-induced uterine cytosolic CTP levels.