Factor requirements for the tritin inactivation of animal cell ribosomes.
Wheat germ contains (tritin) that efficiently inhibits protein synthesis in cell-free extracts from animal cells but not from wheat germ. Tritin has been purified to apparent homogeneity and shown to block enzymatically polypeptide chain elongation. We have extended these studies to examine more closely then mechanism of tritin inactivation of animal cell ribosomes. Here we provide evidence suggesting that ATP and tRNA, previously though to be tritin co-factors, function in the inhibition by altering the conformation of the ribosome to a form susceptible to tritin attack. Tritin treatment does not inhibit the binding of aminoacyl-tRNA to ribosomes, but it does partially reduce the ribosome binding of elongation factor 2. Tritin inhibition appears to affect the core ribosome and not ribosome-associated factors. However, the core ribosome itself is not a suitable substrate for tritin attack; a factor(s) is removed from ribosomes by high salt washing which is required for the tritin-induced inhibition. The ability to be inhibited is restored when ascites cell core ribosomes are supplemented with factors from either ascites cells or wheat germ. In contrast, neither ascites cell factors nor wheat germ factors will promote a significant tritin-induced inhibition of core ribosomes from wheat germ. This indicates that the specificity of tritin inhibition resides primarily at the level of eukaryotic core ribosome.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects; D010445:Peptide Elongation Factors / Q000378:metabolism*; D010940:Plant Proteins / Q000494:pharmacology*; D012343:RNA, Transfer / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D054789:Ribosome Inactivating Proteins, Type 1; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism*; D014908:Triticum
D010445:Peptide Elongation Factors; D010940:Plant Proteins; D012346:RNA, Transfer, Amino Acyl; D054789:Ribosome Inactivating Proteins, Type 1; C031676:tritin protein, Triticum aestivum; D000255:Adenosine Triphosphate; D012343:RNA, Transfer