Journal
EMBO JOURNAL
Volume 26, Issue 9, Pages 2327-2338Publisher
WILEY
DOI: 10.1038/sj.emboj.7601679
Keywords
nonSTOP; ribosome; RNA surveillance; translational repression
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We investigated the fate of aberrant mRNAs lacking in- frame termination codons (called nonSTOP mRNA) in mammalian cells. We found that translation of nonSTOP mRNA was considerably repressed although a corresponding reduction of mRNA was not observed. The repression appears to be post-initiation since (i) repressed nonSTOP mRNAs were associated with polysomes, (ii) translation of IRES-initiated and uncapped nonSTOP mRNA were repressed, and (iii) protein production from nonSTOP mRNA associating with polysomes was significantly reduced when used to program an in vitro run-off translation assay. NonSTOP mRNAs distributed into lighter polysome fractions compared to control mRNAs encoding a stop codon, and a significant amount of heterogeneous polypeptides were produced during in vitro translation of nonSTOP RNAs, suggesting premature termination of ribosomes translating nonSTOP mRNA. Moreover, a run-off translation assay using hippuristanol and RNAse protection assays suggested the presence of a ribosome stalled at the 30 end of nonSTOP mRNAs. Taken together, these data indicate that ribosome stalling at the 30 end of nonSTOP mRNAs can block translation by preventing upstream translation events.
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