Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachussetts, United States of America.
PLoS One. 2012;7(11):e49490. doi: 10.1371/journal.pone.0049490. Epub 2012 Nov 16.
The nonsense mediated decay (NMD) pathway degrades mRNAs bearing premature translation termination codons. In mammals, SMG-8 has been implicated in the NMD pathway, in part by its association with SMG-1 kinase. Here we use four independent assays to show that C. elegans smg-8 is not required to degrade nonsense-containing mRNAs. We examine the genetic requirement for smg-8 to destabilize the endogenous, natural NMD targets produced by alternative splicing of rpl-7a and rpl-12. We test smg-8 for degradation of the endogenous, NMD target generated by unc-54(r293), which lacks a normal polyadenylation site. We probe the effect of smg-8 on the exogenous NMD target produced by myo-3::GFP, which carries a long 3' untranslated region that destabilizes mRNAs. None of these known NMD targets is influenced by smg-8 mutations. In addition, smg-8 animals lack classical Smg mutant phenotypes such as a reduced brood size or abnormal vulva. We conclude that smg-8 is unlikely to encode a component critical for NMD.
无意义介导的衰变(NMD)途径降解带有过早翻译终止密码子的 mRNA。在哺乳动物中,SMG-8 已被牵连到 NMD 途径中,部分原因是它与 SMG-1 激酶有关。在这里,我们使用四种独立的测定方法表明,秀丽隐杆线虫 smg-8 并不需要降解含有无意义的 mRNA。我们检查了 smg-8 对通过 rpl-7a 和 rpl-12 的选择性剪接产生的内源性、天然 NMD 靶标进行不稳定的遗传要求。我们测试了 smg-8 对内源性 NMD 靶标的降解作用,该靶标由 unc-54(r293)产生,它缺乏正常的 poly(A) 位点。我们探测了 smg-8 对由 myo-3::GFP 产生的外源性 NMD 靶标的影响,该 GFP 携带一个长的 3'非翻译区,使 mRNA 不稳定。这些已知的 NMD 靶标都不受 smg-8 突变的影响。此外,smg-8 动物缺乏经典的 Smg 突变表型,例如繁殖力降低或外阴异常。我们得出结论,smg-8 不太可能编码对 NMD 至关重要的成分。