Turnover of the poly(A) moiety of mRNA in wheat-germ extract

The synthesis and degradation of poly(A) were examined in wheat germ lysates under conditions used for protein synthesis. The lysate contained both poly(A)-polymerizing and poly(A)-hydrolytic activities. The synthetic activity was dependent on the presence of either poly(A) or polyadenylated mRNA as primer. Concurrent with the synthesis of poly(A), radioactivity was released from labeled poly(A) and from the poly(A) region of mRNA. Both poly(A) synthesis and hydrolysis were independent of Mn2+ and could proceed in the presence of Mg2+, the major divalent metal ion required for protein synthesis. The synthetic activity was favoured at high (1 mM) ATP concentration, whereas the hydrolytic activity was maximal in the absence of ATP or at low (0.1 mM) ATP concentration. These data indicate that the steady-state length of the poly(A) moeity of mRNAs in the cytoplasm may be regulated by the levels of ATP. Translation products of polyadenylated mRNAs isolated from myeloma cells before and after partial deadenylation were separated by dodecylsulfate/polyacrylamide gel electrophoresis. Shortening of the poly(A) moiety did not alter the relative amounts of the peptides synthesized, indicating that this process does not involve a preferential breakdown of certain species of mRNA.