Control of 4-8S RNA transcription at the midblastula transition in Xenopus laevis embryos.

Abstract

Transcription of Xenopus laevis U1 snRNA genes is subject to a precise program with respect both to the timing of activation at the midblastula transition (MBT) and to the relative levels of the two embryonic U1 RNAs (xU1b1 and b2) that are made. Here, we demonstrate that exogenous xU1b genes injected into developing X. laevis embryos come under the same controls as the endogenous genes. Injected U1 genes, unlike exogenous RNA polymerase III genes, remain quiescent until MBT and their activation at MBT requires protein synthesis during the early cleavage stages. Significantly, the onset of 4-8S RNA transcription occurs at the normal time, even when the DNA content of the embryo has been increased by injection of exogenous DNA or reduced through cleavage arrest, indicating that transcriptional activation at MBT is independent of the ratio of DNA (nucleus) to cytoplasm. In cleavage-arrested (coenocytic) embryos, the reduced level of DNA at MBT results both in a decrease in snRNA and tRNA synthesis (reflecting the lower gene dosage) and in a prolonged synthesis of large amounts of unusual RNA polymerase III transcripts, OAX RNAs. In normally cleaving embryos, small amounts of these unstable OAX RNAs (encoded by satellite I DNA) are synthesized only briefly at MBT. Our demonstration that RNA and DNA metabolism is aberrant in cleavage-arrested embryos requires reevaluation of previous experiments on transcriptional activation that utilized such coenocytic embryos.