Evidence for the regulation of protein synthesis by a wheat germ phosphoprotein factor.
A 48-kilodalton phosphoprotein, termed T-protein or pT, isolated from wheat germ and purified to homogeneity is found to inhibit the translation of tobacco mosaic virus (TMV) RNA in both wheat germ and reticulocyte lysates. The translation of TMV RNA in both systems was inhibited over 80% by 8 microM pT. There was no evidence to indicate that the reticulocyte lysate also contained a pT-like protein. pT was rapidly phosphorylated in the wheat germ and reticulocyte lysates. Although the relationship between pT phosphorylation and inhibition of protein synthesis is not known, there is evidence to indicate that complete phosphorylation of pT is not required for inhibition. Furthermore, no significant differences in the kinetics of inhibition of protein synthesis between prephosphorylated and unmodified pT were observed. Investigation of the mechanism of inhibition indicated that neither the aminoacylation of tRNA nor the elongation of nascent polypeptide chains was affected by pT. On the other hand, pT was found to prevent the formation of the 80S initiation complex. This action of pT was not due to the binding of pT to the ribosomes. However, the effect of pT was found to vary with the concentrations and types of mRNA used in the translational system. These results suggest that pT may interact with specific region(s) of the mRNA and prevent its translation. Alternatively, pT could block the translation of mRNA by binding to one or more of the initiation factors that interact with mRNA to facilitate mRNA binding to the 43S preinitiation complex.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Biological Chemistry, University of Illinois at Chicago, College of Medicine 60612.
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D000818:Animals; D002474:Cell-Free System; D006706:Homeostasis; D007700:Kinetics; D010750:Phosphoproteins / Q000302:isolation & purification / Q000494:pharmacology*; D010940:Plant Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects; D011494:Protein Kinases / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D014027:Tobacco Mosaic Virus / Q000187:drug effects / Q000235:genetics; D014908:Triticum
D010750:Phosphoproteins; D010940:Plant Proteins; D012367:RNA, Viral; C034921:T-substrate protein, Triticum aestivum; D011494:Protein Kinases