Masking and unmasking maternal mRNA. The role of polyadenylation, transcription, splicing, and nuclear history.
We establish that masked mRNAs synthesized from exogenous plasmid templates microinjected into the nuclei of Xenopus oocytes are translationally activated (unmasked) on oocyte maturation concomitant with polyadenylation. Synthetic mRNA injected into the cytoplasm of the oocyte is translated over an order of magnitude more efficiently than is the cognate mRNA synthesized in vivo. Both mRNA synthesized in vivo and mRNA microinjected into the oocyte cytoplasm require a cytoplasmic polyadenylation element in the 3'-untranslated region to activate translation on maturation. Although polyadenylation upon oocyte maturation can relieve the translational repression of mRNA synthesized in vivo, the excision of an intron within the nucleus does not relieve repression. We suggest that the translational repression coupled to the transcription process will more effectively repress inappropriate gene expression in the oocyte and offer the potential to achieve a wider range of gene regulation.
Meric
F
F
Laboratory of Molecular Embryology, NICHD, National Institutes of Health, Bethesda, Maryland 20892-5431, USA. awlme@helix.nih.gov
Searfoss
A M
AM
Wormington
M
M
Wolffe
A P
AP
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D009865:Oocytes / Q000378:metabolism*; D009866:Oogenesis; D011061:Poly A / Q000378:metabolism; D014176:Protein Biosynthesis; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism*; D014158:Transcription, Genetic; D014982:Xenopus laevis
D012333:RNA, Messenger; D011061:Poly A