Romanov Georgy A, Sukhoverov Victor S
Belozersky Institute of Physicochemical Biology, Moscow State University, Moscow, 119991, Russia.
Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, Botanicheskaya 35, Moscow, 127276, Russia.
Mol Genet Genomics. 2017 Oct;292(5):1013-1026. doi: 10.1007/s00438-017-1328-y. Epub 2017 May 18.
Methylation of cytosine residues in DNA of higher eukaryotes, including humans, creates "hot spots" of C→T transitions in the genome. The predominantly methylated sequence in mammalian DNAs is CG (CpG). Among CG-containing codons, CGA codons for arginine are unique due to their ability to create stop codons TGA (UGA in mRNA) upon epigenetic-mediated mutation. As such nonsense mutations can have a strong adverse effect on the cell and organism, we have performed a study, on the example of human genes, aimed to characterise the anticipated effects of epigenetic-mediated nonsense mutations CGA→TGA in somatic cells. It is commonly accepted that premature termination codons (PTCs) lead to the biosynthesis of truncated and usually inactive proteins. In addition, transcripts with PTC can be destroyed by a nonsense-mediated mRNA decay (NMD) machinery. We have considered the cell potentialities (gene families, diploidy, and alternative splicing) to overcome the worst consequences of nonsense mutation. As a special case, in the biosynthesis of a particular group of proteins called selenoproteins, the mutation CGA→UGA would not lead to the premature translation termination and NMD but rather to the insertion of selenocysteine or cysteine instead of former arginine. The finding of SECIS (Sel insertion sequence)-like structures in a variety of mRNAs allowed us to postulate the existence of facultative selenoproteins, whose biosynthesis might be coupled with the redefinition of premature UGA stop codons arising upon mutations, as in the case of "classic" selenoproteins. Nevertheless, a detailed structural analysis of 165 transcripts has shown that roughly 80-90% of functional human mRNAs are potential substrates for NMD upon the PTC emergence. A hypothesis was put forward highlighting a role of arginine CGA codons together with glutamine CAA and CAG codons in the control of mRNA quality and life span. According to this hypothesis, the conversion of the ribonucleic codons CGA, CAA, or CAG into stop codons UGA, UAA or UAG owing to spontaneous or enzymatic cytosine deamination might serve as a trigger for the transcript destruction by NMD (C→U control). Thus, the consequences of epigenetic-mediated nonsense mutations are diverse and may largely depend on the structure of the transcript (CGA codon position, the presence and position of introns and SECIS elements, and splicing potential) of the cognate gene. However, this diversity and the presumable role of CGA codons in performing the everyday function by controlling whether genes are expressed correctly do not exclude their long-term role as limiters of the cell and organism life span. Thus, the presumable role of CGA codons in genome functioning and stability opens new perspectives to influence aging and concomitant deceases by codon editing.
高等真核生物(包括人类)DNA中胞嘧啶残基的甲基化在基因组中产生了C→T转换的“热点”。哺乳动物DNA中主要的甲基化序列是CG(CpG)。在含CG的密码子中,精氨酸的CGA密码子很独特,因为在表观遗传介导的突变后,它们能够产生终止密码子TGA(mRNA中的UGA)。由于这种无义突变会对细胞和生物体产生强烈的不利影响,我们以人类基因为例进行了一项研究,旨在表征表观遗传介导的体细胞中无义突变CGA→TGA的预期影响。人们普遍认为,过早终止密码子(PTC)会导致截短且通常无活性的蛋白质的生物合成。此外,带有PTC的转录本可能会被无义介导的mRNA降解(NMD)机制破坏。我们考虑了细胞的潜力(基因家族、二倍体和可变剪接),以克服无义突变的最坏后果。作为一个特殊情况,在一类称为硒蛋白的特定蛋白质的生物合成中,突变CGA→UGA不会导致过早的翻译终止和NMD,而是导致硒代半胱氨酸或半胱氨酸取代原来的精氨酸插入。在多种mRNA中发现类似SECIS(硒插入序列)的结构使我们推测存在兼性硒蛋白,其生物合成可能与突变后产生的过早UGA终止密码子的重新定义有关,就像“经典”硒蛋白的情况一样。然而,对165个转录本的详细结构分析表明,大约80 - 90%的功能性人类mRNA在出现PTC时是NMD的潜在底物。有人提出了一个假说,强调精氨酸CGA密码子与谷氨酰胺CAA和CAG密码子在控制mRNA质量和寿命方面的作用。根据这个假说,由于自发或酶促的胞嘧啶脱氨作用,核糖核酸密码子CGA、CAA或CAG转换为终止密码子UGA、UAA或UAG可能会触发NMD对转录本的破坏(C→U控制)。因此,表观遗传介导的无义突变的后果是多样的,并且可能在很大程度上取决于同源基因转录本的结构(CGA密码子位置、内含子和SECIS元件的存在及位置以及剪接潜力)。然而,这种多样性以及CGA密码子在通过控制基因是否正确表达来执行日常功能中的假定作用并不排除它们作为细胞和生物体寿命限制因素的长期作用。因此,CGA密码子在基因组功能和稳定性中的假定作用为通过密码子编辑影响衰老和伴随疾病开辟了新的前景。