Cali B M, Anderson P
Department of Genetics, University of Wisconsin, Madison 53706, USA.
Mol Gen Genet. 1998 Nov;260(2-3):176-84. doi: 10.1007/s004380050883.
Nonsense mutant mRNAs are unstable in all eucaryotes tested, a phenomenon termed nonsense-mediated mRNA decay (NMD) or mRNA surveillance. Functions of the seven smg genes are required for mRNA surveillance in Caenorhabditis elegans. In Smg(+) genetic backgrounds, nonsense-mutant mRNAs are unstable, while in Smg(-) backgrounds such mRNAs are stable. Previous work has demonstrated that the elevated level of nonsense-mutant mRNAs in Smg(-) animals can influence the phenotypic effects of heterozygous nonsense mutations. Certain nonsense alleles of a muscle myosin heavy chain gene are recessive in Smg(+) backgrounds but strongly dominant in Smg(-) backgrounds. Such alleles probably express disruptive myosin polypeptide fragments whose abundance is elevated in smg mutants due to elevation of mRNA levels. We report here that mutations in a variety of C. elegans genes are strongly dominant in Smg(-), but recessive or only weakly dominant in Smg(+) backgrounds. We isolated 32 dominant visible mutations in a Smg(-) genetic background and tested whether their dominance requires a functional NMD system. The dominance of 21 of these mutations is influenced by NMD. We demonstrate, furthermore, that in the case of myosin, the dominant-negative effects of nonsense alleles are likely to be due to expression of N-terminal nonsense-fragment polypeptides, not to mistranslation of the nonsense codons. mRNA surveillance, therefore, may mitigate potentially deleterious effects of many heterozygous germline and somatic nonsense or frame-shift mutations. We also provide evidence that smg-6, a gene previously identified as being required for NMD, performs essential function(s) in addition to its role in NMD.
在所有被检测的真核生物中,无义突变的mRNA都是不稳定的,这种现象被称为无义介导的mRNA降解(NMD)或mRNA监测。秀丽隐杆线虫中mRNA监测需要七个smg基因发挥作用。在Smg(+)遗传背景下,无义突变的mRNA是不稳定的,而在Smg(-)背景下,此类mRNA是稳定的。先前的研究表明,Smg(-)动物中无义突变mRNA水平的升高会影响杂合无义突变的表型效应。肌肉肌球蛋白重链基因的某些无义等位基因在Smg(+)背景下是隐性的,但在Smg(-)背景下是强显性的。此类等位基因可能表达具有破坏作用的肌球蛋白多肽片段,由于mRNA水平升高,其丰度在smg突变体中也会升高。我们在此报告,秀丽隐杆线虫多种基因中的突变在Smg(-)背景下是强显性的,但在Smg(+)背景下是隐性的或仅为弱显性。我们在Smg(-)遗传背景中分离出32个显性可见突变,并测试了它们的显性是否需要一个功能性的NMD系统。其中21个突变的显性受NMD影响。此外,我们证明,就肌球蛋白而言,无义等位基因的显性负效应可能是由于N端无义片段多肽的表达,而不是由于无义密码子的错译。因此,mRNA监测可能会减轻许多杂合种系和体细胞无义或移码突变的潜在有害影响。我们还提供证据表明,smg-6这个先前被确定为NMD所需的基因,除了在NMD中的作用外,还执行其他重要功能。