The translational regulation of maternal mRNAs is the primary mechanism by which stage‐specific programs of protein synthesis are executed during early development. Translation of a variety of maternal mRNAs requires either the maintenance or cytoplasmic elongation of a 3′ poly(A) tail. Conversely, deadenylation results in translational inactivation. Although its precise function remains to be elucidated, the highly conserved poly(A) binding protein I (PABP) mediates poly(A)‐dependent events in translation initiation and mRNA stability. Xenopus oocytes contain less than one PABP per poly(A) binding site suggesting that the translation of maternal mRNAs could be either limited by or independent of PABP. In this report, we have analyzed the effects of overexpressing PABP on the regulation of mRNAs during Xenopus oocyte maturation. Increased levels of PABP prevent the maturation‐specific deadenylation and translational inactivation of maternal mRNAS that lack cytoplasmic polyadenylation elements. Overexpression of PABP does not interfere with maturation‐specific polyadenylation, but reduces the recruitment of some mRNAs onto polysomes. Deletion of the C‐terminal basic region and a single RNP motif from PABP significantly reduces both its binding to polyadenylated RNA in vivo and its ability to prevent deadenylation. In contrast to a yeast PABP‐dependent poly(A) nuclease, PABP inhibits Xenopus oocyte deadenylase in vitro. These results indicate that maturation‐specific deadenylation in Xenopus oocytes is facilitated by a low level of PABP consistent with a primary function for PABP to confer poly(A) stability.

The 3 poly(A) structure improves translation of a eukaryotic mRNA by 50-fold in vivo. This enhancement has been suggested to be due to an interaction of the poly(A) binding protein, Pab1p, with eukaryotic translation initiation factor 4G (eIF4G). However, we find that mutation of eIF4G eliminating its interaction with Pab1p does not diminish the preference for poly(A) ؉ mRNA in vivo, indicating another role for poly(A). We show that either the absence of Fun12p (eIF5B), or a defect in eIF5, proteins involved in 60S ribosomal subunit joining, specifically reduces the translation of poly(A) ؉ mRNA, suggesting that poly(A) may have a role in promoting the joining step. Deletion of two nonessential putative RNA helicases (genes SKI2 and SLH1) makes poly(A) dispensable for translation. However, in the absence of Fun12p, eliminating Ski2p and Slh1p shows little enhancement of expression of non-poly(A) mRNA. This suggests that Ski2p and Slh1p block translation of non-poly(A) mRNA by an effect on Fun12p, possibly by affecting 60S subunit joining.The 5Ј cap (7-methyl-GpppG. . .) and 3Ј poly(A) structures of eukaryotic mRNAs are both critically important for translation, for mRNA transport from the nucleus, and for mRNA stability in both the nucleus and the cytoplasm. The in vivo requirement for translation for the 3Ј poly(A) structure, estimated by electroporation of mRNAs into living cells, is about 50-fold, while that for the 5Ј cap is about 20-fold (11, 12).The effects of the 5Ј cap on translation are mediated by the cap-binding protein, eukaryotic translation initiation factor 4E (eIF4E) (Cdc33p), which associates with eIF4G in the eIF4F complex to promote binding of the mRNA to the 40S ribosomal subunit (through eIF3). A 43S preinitiation complex consisting of the 40S subunit, eIF3, and eIF2-GTP-MettRNA iMet is thought to bind to an mRNA mediated by interaction between eIF3 and eIF4G. This complex scans from the cap at the 5Ј end of the mRNA to the first AUG. There, with the help of eIF5 (Tif5p) and eIF5B (Fun12p), 60S subunit joining occurs and translation begins (27; reviewed in references 9 and 37). 60S subunit joining requires both eIF5 and eIF5B. While eIF5 promotes GTP hydrolysis by eIF2, enabling release of the initiation factors from the 40S subunit, eIF5B has its own GTP binding activity and hydrolyzes GTP in a ribosome-dependent reaction (27).The role of the 3Ј poly(A) structure in mRNA translation is not yet completely clear. The poly(A) binding protein (Pab1p) is believed to mediate many of the effects of the 3Ј poly(A) structure (reviewed in reference 34). Because the poly(A) tail apparently has roles in nuclear processes as well as cytoplasmic events (22), dissecting these mechanisms is difficult. The PAB1 gene is essential, indicating that at least one of the functions of Pab1p is critical to the cell.

In wild-type cells, the 3' poly(A) structure is necessary for translation of mRNA and for mRNA stability. The superkiller 2 (ski2), ski3, ski6, ski7, and ski8 mutations enhance the expression of the poly(A)(-) mRNAs of yeast RNA viruses. Ski2p is a DEVH-box RNA helicase and Slh1p resembles Ski2p. Both repress L-A double-stranded RNA (dsRNA) virus copy number, further suggesting that their functions may overlap. We find that slh1Delta ski2Delta double mutants are healthy (in the absence of viruses) and show normal rates of turnover of several cellular mRNAs. The slh1Delta ski2Delta strains translate electroporated nonpoly(A) mRNA with the same kinetics as polyA(+) mRNA. Thus, the translation apparatus is inherently capable of efficiently using nonpoly(A) mRNA even in the presence of normal amounts of competing poly(A)(+) mRNA, but is normally prevented from doing so by the combined action of the nonessential proteins Ski2p and Slh1p.