Dexamethasone stimulates ribosomal protein L32 gene transcription in rat myoblasts.

Incubation of rat L6 myoblasts for 24 h with 10(-7) M dexamethasone, a glucocorticoid analogue, resulted in a 2.5-fold increase in the rate of ribosomal protein L32 (rpL32) gene transcription with a corresponding increase in the level of rpL32 mRNA. The increased rate of transcription was accompanied by a dramatic enhancement in binding of the delta, but not beta and gamma, factors to the rpL32 gene promoter as measured by gel mobility shift assays. This increased binding reflects a change in the activity of the delta factor since its level is unchanged by dexamethasone treatment. The presence of the glucocorticoid analogue RU38486 reversed the stimulating effect of dexamethasone on rpL32 gene transcription and binding of the delta factor to the delta element. These results suggest that the mechanism which enhances rpL32 gene transcription in dexamethasone-treated rat L6 myoblasts involves glucocorticoid-receptor mediated changes in the activity of the delta factor.

Altered subcellular distribution of U3 snRNA in response to serum in mouse fibroblasts.

To extend our understanding of the mechanisms regulating ribosome biosynthesis during changes in cellular growth rate, the expression and subcellular distribution of U3 snRNA and one of its associated proteins, fibrillarin, were examined in mouse 3T6 fibroblasts. Altering serum concentrations produces changes in the ribosome content of the cell as reflected by total RNA levels. When exponentially growing 3T6 cells are induced to become quiescent by serum starvation, a significant downshift in U3 snRNA gene transcription occurs in parallel to a decrease in pre-rRNA synthesis. Serum stimulation results in an increase in the rate of synthesis of both U3 snRNA and pre-rRNA. However, U3 snRNA synthesis lags behind that of pre-rRNA. Furthermore, in serum-starved fibroblasts, a significant portion of the total cellular U3 snRNA appears in the cytoplasm. Following serum stimulation, a redistribution occurs and U3 snRNA is localized predominantly in the nucleolus at a level similar to that observed in exponentially growing cells. This redistribution is inhibited when RNA or protein synthesis is repressed in serum-stimulated fibroblasts by actinomycin D or cycloheximide. In contrast, the level and subcellular distribution of fibrillarin remain unchanged during serum starvation. These results suggest that during changes in ribosome production, distinct pools of U3 snRNPs exist within the cell.

Regulation of U3 snRNA expression during myoblast differentiation.

Differentiation of proliferating rat L6 myoblasts to syncytial multinucleated myotubes results in a significant downshift in the rate of U3 snRNA gene transcription, paralleling the decrease in rRNA synthesis previously documented. Coordinate production of U3 snRNA and rRNA during the differentiation process adds further support for a role of U3 snRNA in ribosome biogenesis. Despite the dramatic decrease in U3 snRNA transcription during differentiation, a corresponding drop in the cellular level of U3 snRNA does not occur. In myotubes, the amount of U3 snRNA is regulated at the post-transcriptional level in which there is a significant accumulation of U3 snRNA in the cytoplasm of myotubes. This intracellular redistribution of U3 snRNA may significantly affect the entire process of rRNA maturation or result from the decrease in ribosome production accompanying terminal differentiation of myoblasts.