ABSTRACT
To study the regulation of AUUUA-mediated RNA deadenylation and destabilization during Xenopus early development, we microinjected chimeric mRNAs containing Xenopus or mammalian 3′ untranslated region (3′-UTR) sequences intoXenopus oocytes, mature eggs, or fertilized embryos. We found that the AU-rich elements (ARE) of Xenopusc-myc II and the human granulocyte-macrophage colony-stimulating factor gene (GMCSF) both direct deadenylation of chimeric mRNAs in an AUUUA-dependent manner. In the case of the Xenopus c-myc II ARE, mutation of a single AUUUA within an absolutely conserved 11-nucleotide region in c-myc 3′-UTRs prevents ARE-mediated deadenylation. AUUUA-specific deadenylation appears to be developmentally regulated: low deadenylation activity is observed in the oocyte, whereas rapid deadenylation occurs following egg activation or fertilization. Deadenylation results in the accumulation of stable deadenylated RNAs that become degraded only following mid-blastula transition. We conclude that ARE-mediated mRNA deadenylation can be uncoupled from ARE-mediated mRNA decay and that AUUUAs directly signal deadenylation during Xenopus early development.
ACKNOWLEDGMENTS
We thank B. DeDecker, D. Enke, C. Fan, M. Frilander, T. McConnell, L. Scharl, and L. Weinstein for many helpful suggestions and careful reading of the manuscript and B. Egan and S. Baserga for plasmids.
This work was supported by grant CA16038 from the National Institutes of Health.