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- W3147673668 abstract "Nonsense-mediated mRNA decay( NMD) acts as an important supervision and quality control mechanism regulating the elimination of mRNA containing premature translation termination codon( PTC). PTC is produced by nonsense,miss-splicing,frame-shift mutations. NMD pathway-mediated mRNA degradation will prevent the deleterious damage caused by improperly truncated proteins. Manyaberrant mRNAs containing PTC could escape from the NMD pathway,but the mechanism remains unclear. In this study,we constructed a mini-RB1( retinoblastoma gene 1,RB1) fragment,which included c DNA of exon 1 ~ 14,DNA of intron 14-exon 15-intron 15 and c DNA of exon 16 ~ 27,and inserted it into mammalian expression vector pc DNA3. 1(-). Three nonsense mutants of mini-RB1 were generated by introducing W99 X,G310X and R467 X into mini-RB1 gene,individually,based on Human Genome Mutation Database( HGMD). The real-time q PCR was used to analyze the mRNA level of the mini-RB1 before and after treatment with actinomycin D,an inhibitor of transcription,or cycloheximide,an inhibitor of NMD pathway. No significant difference of mRNA levels of mini-RB1( W99X) mutants was observed with treatment of the either of the two drugs. Also,comparable mRNA levels were observed between the wild-type mini-RB1 and mini-RB1 mutants. All these facts suggest that the RB1( W99X)might be capable of escaping from the NMD pathway. Furthermore,Western blotting indicated that the translation of mini-RB1( W99X) was re-initiated downstream of W99 X and the resultant N-terminus truncated proteins of RB1 might lead to the escape of the nonsense mRNA from NMD. Our results highlighted the possibility that translation re-initiation of the nonsense W99 X mutation down-stream might be the major mechanism responsible for the NMD escape of RB1 mutants." @default.
- W3147673668 created "2021-04-13" @default.
- W3147673668 creator A5033553707 @default.
- W3147673668 date "2015-01-01" @default.
- W3147673668 modified "2023-09-23" @default.
- W3147673668 title "Translation Re-initiation of PTC Downstream Induced the Escape of Nonsense mRNA from NMD" @default.
- W3147673668 hasPublicationYear "2015" @default.
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