The effects of glucose and cyclic GMP on RNA synthesis and nuclear morphology in starved rats.

Feeding rats in diet high in glucose has been demonstrated to inhibit the induction of many enzymes, block the action of glucocorticoids, and, in general, appears to result in decreased cyclic AMP activity. We found that glucose feeding depresses both messenger RNA (mRNA) and non-mRNA synthesis. Electron microscopic examination of the nucleus revealed that glucose feeding decreases the granular component of liver cell nucleoli. It only slightly decreases liver cyclic AMP levels, but produces a sixfold elevation in levels of the cyclic AMP antagonist, cyclic GMP. Administration of bromocyclic GMP, like glucose feeding, depresses mRNA synthesis, but does not simulate the effect of the carbohydrate on nuclear morphology. In addition, glucose feeding halves liver inorganic phosphate and triples ATP levels. Phosphorylation of nuclear proteins, however, remains unaltered. Despite the antagonism between glucose feeding and glucocorticoid activity, the former compound did not change the binding of dexamethasone to liver nuclei.     

Methylation of DNA guanine via the 1-carbon pool in dimethylnitrosamine-treated rats

Treatment of rats with radioactive methionine and nonradioactive dimethylnitrosamine resulted in the formation of radioactive 7-methylguanine in rat-liver DNA. By comparing the specific activity of administered [14C-Me]-dimethylnitrosamine to the specific activity of isolated 7-methylguanine it was determined that following 20 mg/kg dimethylnitrosamine DNA methylation via the 1-carbon pool may account for up to 30% of the total 7-methylguanine formed.     

Ultraviolet light-induced inhibition of small nuclear RNA synthesis

Two apparently distinct types of inhibition of the synthesis of U1, U2, U3, U4, and U5 small nuclear RNA, induced by ultraviolet (UV) radiation, have been described before: immediate and delayed. Our present observation can be summarized as follows: a) neither the immediate nor the delayed inhibition appear to be mediated by the formation of cyclobutane pyrimidine dimers, since they were not prevented by photoreactivating light, in ICR 2A frog cells; b) the inhibition of U1 RNA synthesis, monitored in HeLA cells within the first few minutes after irradiation, extrapolated to a substantial suppression at time zero of postirradiation cell incubation, providing further support for the proposal that the immediate inhibition is a reaction separate from the delayed UV light-induced inhibition of U1 RNA synthesis; c) the transition from the pattern of the immediate inhibition to that of the delayed inhibition (disappearance of the UV-resistant fraction of U1 RNA synthesis and increased rate of inhibition) occurred gradually, without an apparent threshold, within the first 2 hr of incubation after irradiation; and d) the incident UV dose that resulted in a 37% level of residual U1 RNA synthesis (D37) during the delayed inhibition was about 7 J/m2, with an apparent UV dose threshold, and was about 60 J/m2 for the immediate inhibition.     

Liver and kidney cortex gluconeogenesis from L-alanine in fed and starved rats

Circulating [14C]glucose 2, 5 and 10 min after intravenous injection of [U-14C]-L-alanine was greater in 24 hr starved than in fed rats. In vitro uptake of [14C]alanine by liver and kidney cortex slices from 24 hr starved and fed rats rose in parallel with increased medium substrate concentration. Formation of [14C]glucose from 1mM [14C]alanine was similar in liver and kidney cortex slices and increased in tissues from 24 hr starved compared with fed rats. With 5 mM [14C]alanine more [14C]glucose was produced by liver than by kidney cortex slices from 24 hr starved rats. Liver slices always produced more [14C]lactate and less [14C]-CO2 from [14C]alanine than kidney cortex slices. It is proposed that under physiological conditions, the kidneys cortex actively participates in glucose production from alanine.     

Altered nuclear RNA transport associated with carcinogen intoxication in rats

The release of nuclear restricted RNA to the cytoplasm in dimethylaminoazobenzene (DAB) and acetylaminofluorene (AAF) induced tumors suggested a basis for the phenotypic expression of chemically induced neoplasms. In this report the increased release of RNA from nuclei $\text{in vitro}$ is identified both following acute intoxication and with prolonged (tumor producing) exposure to a third carcinogen, thioacetamide. This release is independent of ATP added in the cell-free system and consistent with “leaky” nuclei, providing further support for the hypothesis that nuclear RNA transport is involved in chemical carcinogenesis.