Multiple levels of regulation of protein synthesis at fertilization in sea urchin eggs.
Fertilization of sea urchin eggs results in a large stimulation of protein synthesis. This increase in protein synthesis is mediated by the mobilization of stored maternal mRNA (mRNPs) into polysomes, but the details of the molecular mechanisms which regulate this process are not well understood. Using a sea urchin egg cell-free translation system, evidence has been obtained which indicates that the capacity to initiate protein synthesis on new mRNAs is limited. Addition of exogenous mRNAs failed to stimulate overall protein synthesis, whereas supplementing the system with a nuclease-treated reticulocyte lysate, an S-100 supernatant fraction, or purified eIF-2 stimulated nearly twofold. In addition, the levels of 43 S preinitiation complexes containing a 40 S ribosomal subunit and methionyl-tRNA were increased at pH 7.4 compared to pH 6.9, or when reticulocyte S-100 was added. However, other experiments showed clearly that mRNA availability may also regulate translation in the sea urchin egg. Sea urchin lysates only stimulated poorly the nuclease-treated reticulocyte lysate system, and the mRNPs in the sea urchin lysate did not bind to reticulocyte 43 S preinitiation complexes. Since purified sea urchin egg mRNA was active in both assays, the bulk of sea urchin mRNA must be masked in the egg, and remain masked in the in vitro assays. Thus, protein synthesis appears to be regulated at both the level of mRNA availability and the activity of components of the translational machinery.
D000818:Animals; D002118:Calcium / Q000502:physiology; D002474:Cell-Free System; D005306:Fertilization; D005786:Gene Expression Regulation; D006863:Hydrogen-Ion Concentration; D010442:Peptide Chain Initiation, Translational; D010448:Peptide Initiation Factors / Q000502:physiology; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D012617:Sea Urchins / Q000196:embryology*
D010448:Peptide Initiation Factors; D012333:RNA, Messenger; D002118:Calcium