The synthesis of ribosomal proteins S16 and L32 is not autogenously regulated during mouse myoblast differentiation.
A series of mouse myoblast cell lines was constructed that contain 1 to 34 extra copies of either the S16 or the L32 ribosomal protein (r-protein) gene. The metabolism of the S16 and L32 r-proteins and mRNAs was examined in myoblasts and fibers of these cell lines to determine whether the synthesis of these r-proteins is autogenously regulated. The incorporation of extra copies of these r-protein genes into the genome resulted in the accumulation of the corresponding mRNAs to levels that were directly proportional to the gene copy number. The levels of the overproduced mRNAs decreased after the differentiation of mouse myoblasts into fibers in parallel to the decrease in the levels of the endogenous r-protein mRNAs. These results indicate that the synthesis of these r-proteins is not autogenously regulated at the level of transcription, RNA processing, or mRNA stability. To determine whether the synthesis of these r-proteins is regulated at the level of translation, the translational efficiencies of the overproduced mRNAs were inferred from their distribution in polysomal gradients. The translational efficiencies of these overproduced r-protein mRNAs in myoblasts are similar to those of the endogenous r-protein mRNAs. After myoblast differentiation, the translational efficiencies of the overproduced r-protein mRNAs decrease exactly like those of the endogenous r-protein mRNAs. Examination of the synthesis and stability of r-proteins in one of the L32-overproducing cell lines demonstrated that the overproduced L32 r-protein degrades shortly after its synthesis. The synthesis and stability of the other r-proteins were unaffected in this cell line. Thus, the synthesis of S16 and L32 r-proteins is not autogenously regulated at any level in either myoblasts or fibers.
Department of Biology, University of South Carolina, Columbia 29208.
L H
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Bowman
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004247:DNA / Q000235:genetics; D005786:Gene Expression Regulation; D051379:Mice; D009132:Muscles / Q000166:cytology* / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D012269:Ribosomal Proteins / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection
D012333:RNA, Messenger; D012269:Ribosomal Proteins; C041806:ribosomal protein L32; C023509:ribosomal protein S16; D004247:DNA