Génétique des Interactions Macromoléculaires, Institut Pasteur, CNRS UMR3525, 25-28 rue du docteur Roux, 75015 Paris, France.
Production de Protéines Recombinantes et d'Anticorps (PF5), Institut Pasteur, 25-28 rue du docteur Roux, 75015 Paris, France.
Cell Rep. 2014 Feb 27;6(4):593-8. doi: 10.1016/j.celrep.2014.01.025. Epub 2014 Feb 13.
Nonsense-mediated mRNA decay (NMD) destabilizes eukaryotic transcripts with long 3' UTRs. To investigate whether other transcript features affect NMD, we generated yeast strains expressing chromosomal-derived mRNAs with 979 different promoter and open reading frame (ORF) regions and with the same long, destabilizing 3' UTR. We developed a barcode-based DNA microarray strategy to compare the levels of each reporter mRNA in strains with or without active NMD. The size of the coding region had a significant negative effect on NMD efficiency. This effect was not specific to the tested 3' UTR because two other different NMD reporters became less sensitive to NMD when ORF length was increased. Inefficient NMD was not due to a lack of association of Upf1 to long ORF transcripts. In conclusion, in addition to a long 3' UTR, short translation length is an important feature of NMD substrates in yeast.
无意义介导的 mRNA 降解 (NMD) 会使具有长 3'UTR 的真核转录本不稳定。为了研究其他转录本特征是否会影响 NMD，我们构建了表达来自染色体的带有 979 种不同启动子和开放阅读框（ORF）区且具有相同长的、不稳定 3'UTR 的 mRNA 的酵母菌株。我们开发了一种基于条形码的 DNA 微阵列策略来比较有或没有活性 NMD 的菌株中每个报告 mRNA 的水平。编码区的大小对 NMD 效率有显著的负向影响。这种影响不是针对所测试的 3'UTR 的，因为当 ORF 长度增加时，另外两个不同的 NMD 报告基因对 NMD 的敏感性降低。NMD 效率低下不是由于 Upf1 与长 ORF 转录本的结合不足所致。总之，除了长 3'UTR 之外，短的翻译长度也是酵母中 NMD 底物的一个重要特征。
