Destabilization of tubulin mRNA during heat shock in Tetrahymena pyriformis.

Abstract

The regulation of tubulin gene expression was studied in Tetrahymena pyriformis cells during heat shock (shift from 28 degrees C to 34 degrees C). Fluorograms of two-dimensional gels of radiolabelled proteins synthesized during thermal stress revealed that tubulin synthesis is highly repressed when compared with that of exponentially growing cells. The variation in the levels of alpha and beta-tubulin mRNAs was analyzed by Northern-blot hybridization using homologous genomic probes (alpha TT and beta TT1). The results obtained show that heat shock induces a drastic and coordinate reduction in the amount of alpha and beta-tubulin mRNAs isolated from polysomes. This decrease is not due to a shift from the polysomes to the post-polysomal fraction because it was also observed when total cytoplasmic mRNAs were analyzed. Run-on transcription experiments were performed in order to examine whether repression of transcription in heat-shocked cells could explain that reduction. The results obtained show that the apparent rates of tubulin gene transcription are not significantly modified, but on the contrary increase slightly in cells heat-shocked for 15 min and 30 min. The effects of inhibitors of protein synthesis, cycloheximide and pactamycin, on the destabilization of tubulin mRNAs were tested in heat-shocked Tetrahymena cells. Our results revealed that in the presence of these inhibitors, tubulin mRNAs become more stable thus suggesting that an induced factor may be involved in the degradation of alpha and beta-tubulin mRNAs during heat shock.