Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States of America.
PLoS One. 2012;7(10):e47085. doi: 10.1371/journal.pone.0047085. Epub 2012 Oct 11.
Expanded trinucleotide repeat sequences are the cause of several inherited neurodegenerative diseases. Disease pathogenesis is correlated with several features of somatic instability of these sequences, including further large expansions in postmitotic tissues. The presence of somatic expansions in postmitotic tissues is consistent with DNA repair being a major determinant of somatic instability. Indeed, proteins in the mismatch repair (MMR) pathway are required for instability of the expanded (CAG·CTG)(n) sequence, likely via recognition of intrastrand hairpins by MutSβ. It is not clear if or how MMR would affect instability of disease-causing expanded trinucleotide repeat sequences that adopt secondary structures other than hairpins, such as the triplex/R-loop forming (GAA·TTC)(n) sequence that causes Friedreich ataxia. We analyzed somatic instability in transgenic mice that carry an expanded (GAA·TTC)(n) sequence in the context of the human FXN locus and lack the individual MMR proteins Msh2, Msh6 or Pms2. The absence of Msh2 or Msh6 resulted in a dramatic reduction in somatic mutations, indicating that mammalian MMR promotes instability of the (GAA·TTC)(n) sequence via MutSα. The absence of Pms2 resulted in increased accumulation of large expansions in the nervous system (cerebellum, cerebrum, and dorsal root ganglia) but not in non-neuronal tissues (heart and kidney), without affecting the prevalence of contractions. Pms2 suppressed large expansions specifically in tissues showing MutSα-dependent somatic instability, suggesting that they may act on the same lesion or structure associated with the expanded (GAA·TTC)(n) sequence. We conclude that Pms2 specifically suppresses large expansions of a pathogenic trinucleotide repeat sequence in neuronal tissues, possibly acting independently of the canonical MMR pathway.
三核苷酸重复序列扩增是几种遗传性神经退行性疾病的病因。疾病的发病机制与这些序列的体细胞不稳定性的几个特征相关,包括在后有丝分裂组织中进一步的大扩展。在后有丝分裂组织中存在体细胞扩展与 DNA 修复是体细胞不稳定性的主要决定因素一致。事实上,错配修复 (MMR) 途径中的蛋白质对于扩增的 (CAG·CTG)(n) 序列的不稳定性是必需的,可能通过 MutSβ 识别链内发夹。目前尚不清楚 MMR 是否会影响除发夹以外的其他二级结构(如引起弗里德里希共济失调的三链体/R 环形成 (GAA·TTC)(n) 序列)的致病三核苷酸重复序列的不稳定性。我们分析了携带 FXN 基因座中扩增 (GAA·TTC)(n) 序列的转基因小鼠的体细胞不稳定性,这些小鼠缺乏单个 MMR 蛋白 Msh2、Msh6 或 Pms2。Msh2 或 Msh6 的缺失导致体细胞突变明显减少,表明哺乳动物 MMR 通过 MutSα 促进 (GAA·TTC)(n) 序列的不稳定性。Pms2 的缺失导致神经系统(小脑、大脑和背根神经节)中大量扩增的积累增加,但在非神经元组织(心脏和肾脏)中没有影响,而不影响收缩的流行率。Pms2 特异性抑制显示 MutSα 依赖性体细胞不稳定性的组织中的大扩展,表明它们可能作用于与扩增的 (GAA·TTC)(n) 序列相关的相同病变或结构。我们得出结论,Pms2 特异性抑制神经元组织中致病性三核苷酸重复序列的大扩展,可能独立于经典的 MMR 途径。