Multiple levels of regulation of tubulin gene expression during sea urchin embryogenesis

Abstract

The expression of the tubulin genes during embryogenesis of the sea urchin Lytechinus pictus has been analyzed. Single strand tracer excess titrations of alpha- and beta-tubulin mRNA and RNA gel blot hybridizations indicate that tubulin mRNA remains at a constant 1.3 X 10(5) transcripts per embryo during cleavage stages, increases during ciliogenesis shortly before hatching (12 hr PF), declines until midgastrula (30-35 hr PF), and then gradually increases 3-fold to about 6 X 10(5) per pluteus larva (72 hr PF). Tubulin synthesis changes in concert with its mRNA, except that during cleavage the relative rate of tubulin synthesis increases without a corresponding increase in tubulin mRNA abundance. The relative rates of tubulin gene transcription were assayed by a run-on assay in isolated nuclei. The synthesis of alpha- or beta-tubulin RNA results in little supplementation of maternal tubulin RNA during cleavage stages, but the rate increases at least 18-fold during ciliogenesis and then gradually decreases thereafter. The accumulation of tubulin mRNA after gastrulation can be accounted for by an ontogenetic increase in tubulin RNA stability, assayed by actinomycin D chase and RNA gel blot hybridization. The rates of synthesis, stabilities, and abundances of alpha- and beta-tubulin mRNAs were similar, suggesting coordinate regulation. These observations indicate the importance of translational regulation during cleavage, transcriptional regulation during ciliogenesis, and regulation of mRNA stability by the level of unpolymerized tubulin during later development.