Regulation of polyribosome formation and protein synthesis in the uterus. Effect of ovariectomy and administration of oestradiol-17β on the amino acid-incorporation activity in vitro and the cytoplasmic concentration in vivo of polyribosomal preparation

Abstract

1. The hormonal regulation of cytoplasmic protein synthesis in the uterus is described. Polyribosomal preparation from uteri of normal or ovariectomized rats was isolated by procedure 3 and assayed for [14C]leucine-incorporation activity in the cell-free system, as described by Teng & Hamilton (1967). 2. Ovariectomy of normal animals caused, 3 weeks after surgery, a 50–60% increase in the amino acid-incorporation activity in vitro of uterine polyribosomal preparation, but a 90–95% decrease in the cytoplasmic concentration in vivo of the preparation. 3. Administration of 10μg. of oestradiol-17β to ovariectomized rats at zero time caused, 10–12hr. later, a 100% stimulation in amino acid-incorporation activity in vitro of the uterine polyribosomal preparation. From 12hr. to 36hr. after hormone administration, the activity in vitro of the preparation decreased. If a second dose of hormone was administered at 36hr., the activity in vitro of the preparation continued to decrease, and approached at 48hr. and 72hr. the lower activity observed for the preparation from normal animals. The cytoplasmic concentration of polyribosomal preparation increased by 600–700% under these experimental conditions. If a second dose of oestradiol-17β was not administered at 36hr., the initially elevated cytoplasmic concentration of the preparation decreased by 50% from 36hr. to 72hr., and the activity in vitro of the preparation was not fully depressed to the ‘normal’ value. 4. Pretreatment of ovariectomized animals with actinomycin D or cycloheximide abolished 80–90% of the stimulatory effects of hormone treatment on the amino acid-incorporation activity in vitro and cytoplasmic concentration in vivo of uterine polyribosomal preparation. 5. Two major conclusions are drawn from the results reported: that during early oestrogen action new polyribosomes having amino acid-incorporation properties different from those of the old ones appear and accumulate in the cytoplasm of the uterus; and that the regulation of cytoplasmic protein synthesis in the organ by oestrogen is of an indirect nature, with dual effects of the hormone on genetic transcription resulting in turn in a regulation of the rate and amount of genetic translation.