Further studies on the stimulation of protein synthesis in androgen-dependent tissues by testosterone

1. By using centrifugation through a discontinuous sucrose gradient, four microsomal fractions are obtained from the prostate gland. 2. Administration of androgens to castrated rats stimulates protein synthesis in all fractions, particularly in the heavy rough fraction. 3. Androgens also increase the content of protein, RNA and phospholipid in the heavy rough fraction. 4. Time-course experiments in vivo show that androgens induce a rapid increase in the synthesis of ribosomal precursor RNA preceding the synthesis of new microsomal fraction and the increase in protein synthesis.     

Formation of ribothymidine from thymine and ribonucleosides by the cell-free extract of tumors and rat tissues.

Formation of ribothymidine by the ribose exchange reaction between thymine and uridine with the cell-free extract of mouse Ehrlich ascites tumor cells was demonstrated. Since phosphate ions appear to be not required for this reaction, perhaps it proceeds by the mechnism of direct exchange of nucleoside N-ribosyltransferase. The transfer activity was found in the precipitates when the crude extract was fractionated with 30-60% saturated ammonium sulfate. Ribothymidine formation was also demonstrated between thymine and ribonucleosides other than uridine with this tumor extract. Production of ribothymidine from thymine and uridine was detected also by the use of extracts from lung, brain, and regenerating liver of normal rats, and from newborn rats (whole body). An extract of Rhodamine sarcoma exhibited the ribose exchange activity, while that of human gastric cancer did not.     

Further studies on the cell-free synthesis of procollagen-collagen by chick embryo polysomes

Free and membrane bound polysomes were prepared from 8-day-old chick embryos. Both polysome preparations were equally active in protein synthesis but procollagen-collagen synthesis was carried out exclusively by the membrane bound polysomes. The collagenous product was analyzed by DEAE-cellulose chromatography and after hydroxylation with peptidyl proline hydroxylase had a hydroxyproline/proline ratio of 0.77. This suggests that the collagenous product synthesized by the membrane bound polysomes is procollagen.     

Comparative studies on the methionine synthesis in sheep and rat tissues

The important features of the enzymes involved in methionine synthesis in sheep were found to be the low activity of betaine-homocysteine methyltransferase and the high activity of 5-methyltetrahydrofolate-homocysteine methyltransferase. The rate of the methionine synthesis in sheep liver was significantly lower than that in rats due to the low activity of hepatic betaine-homocysteine methyltransferase. The hepatic methionine recycling was stimulated by the addition of betaine in both species. These results indicate that in sheep 5-methyltetrahydrofolate-homocysteine methyltransferase plays a significant role in hepatic methionine synthesis along with betaine-homocysteine methyltransferase. In contrast, in the rat hepatic system methionine synthesis is virtually dependent on betaine-homocysteine methyltransferase.