The phosphorylation of protein S6 modulates the interaction of the 40 S ribosomal subunit with the 5'-untranslated region of a dictyostelium pre-spore-specific mRNA and controls its stability.
AC914 mRNA, a pre-spore-specific mRNA that accumulates only in the post-aggregation stage of development, is transcribed constitutively as shown by nuclear run-off experiments and by fusing its promoter to the luciferase reporter gene. The same mRNA disappears quickly from disaggregated cells. If the 5'-untranslated region (5'UTR) of the constitutively expressed Actin 15 mRNA is substituted for the 5'UTR of AC914 mRNA, this can no longer be destabilized and accumulates both in growing and disaggregated cells. If the 5'UTR of AC914 mRNA is substituted for the 5'UTR of Actin 15 mRNA, the latter accumulates only in aggregated cells. Pactamycin, but not other inhibitors of protein synthesis, prevents AC914 mRNA from being destabilized in disaggregated cells, suggesting a role of 40 S subunits in the destabilization. This has been confirmed by using an in vitro system in which the in vivo stability of different mRNAs is reproduced. A protein kinase A-dependent phosphorylation of ribosomal protein S6 determines whether 40 S subunits are capable or not of destabilizing AC914 mRNA in the in vitro system.
Department of Clinical and Biological Sciences, University of Turin, Ospedale S. Luigi, Orbassano (To), Italy 10043.
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S
Chiaberge
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Cassarino
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G
Mangiarotti
D020121:5' Untranslated Regions; D000818:Animals; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004023:Dictyostelium / Q000378:metabolism*; D010142:Pactamycin / Q000494:pharmacology; D010766:Phosphorylation; D011132:Polyribosomes / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012331:RNA, Fungal / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*; D013172:Spores, Fungal; D014158:Transcription, Genetic
D020121:5' Untranslated Regions; D011500:Protein Synthesis Inhibitors; D012331:RNA, Fungal; D012333:RNA, Messenger; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins; D010142:Pactamycin; D017868:Cyclic AMP-Dependent Protein Kinases