Pelassa Ilaria, Corà Davide, Cesano Federico, Monje Francisco J, Montarolo Pier Giorgio, Fiumara Ferdinando
Department of Neuroscience and.
Center for Molecular Systems Biology, University of Torino, Torino 10123, Italy.
Hum Mol Genet. 2014 Jul 1;23(13):3402-20. doi: 10.1093/hmg/ddu049. Epub 2014 Feb 4.
The expansion of homopolymeric glutamine (polyQ) or alanine (polyA) repeats in certain proteins owing to genetic mutations induces protein aggregation and toxicity, causing at least 18 human diseases. PolyQ and polyA repeats can also associate in the same proteins, but the general extent of their association in proteomes is unknown. Furthermore, the structural mechanisms by which their expansion causes disease are not well understood, and these repeats are generally thought to misfold upon expansion into aggregation-prone β-sheet structures like amyloids. However, recent evidence indicates a critical role for coiled-coil (CC) structures in triggering aggregation and toxicity of polyQ-expanded proteins, raising the possibility that polyA repeats may as well form these structures, by themselves or in association with polyQ. We found through bioinformatics screenings that polyA, polyQ and polyQA repeats have a phylogenetically graded association in human and non-human proteomes and associate/overlap with CC domains. Circular dichroism and cross-linking experiments revealed that polyA repeats can form--alone or with polyQ and polyQA--CC structures that increase in stability with polyA length, forming higher-order multimers and polymers in vitro. Using structure-guided mutagenesis, we studied the relevance of polyA CCs to the in vivo aggregation and toxicity of RUNX2--a polyQ/polyA protein associated with cleidocranial dysplasia upon polyA expansion--and found that the stability of its polyQ/polyA CC controls its aggregation, localization and toxicity. These findings indicate that, like polyQ, polyA repeats form CC structures that can trigger protein aggregation and toxicity upon expansion in human genetic diseases.
由于基因突变,某些蛋白质中同聚谷氨酰胺(polyQ)或丙氨酸(polyA)重复序列的扩增会诱导蛋白质聚集并产生毒性,导致至少18种人类疾病。PolyQ和polyA重复序列也可存在于同一蛋白质中,但它们在蛋白质组中的总体关联程度尚不清楚。此外,其扩增导致疾病的结构机制尚未完全了解,一般认为这些重复序列在扩增后会错误折叠成易于聚集的β-折叠结构,如淀粉样蛋白。然而,最近的证据表明卷曲螺旋(CC)结构在触发polyQ扩增蛋白的聚集和毒性方面起关键作用,这增加了polyA重复序列自身或与polyQ结合也可能形成这些结构的可能性。我们通过生物信息学筛选发现,polyA、polyQ和polyQA重复序列在人类和非人类蛋白质组中具有系统发育分级关联,并与CC结构域相关联/重叠。圆二色性和交联实验表明polyA重复序列可以单独或与polyQ和polyQA一起形成CC结构,其稳定性随polyA长度增加,在体外形成高阶多聚体和聚合物。利用结构导向诱变,我们研究了polyA CC对RUNX2体内聚集和毒性的相关性,RUNX2是一种polyQ/polyA蛋白,在polyA扩增时与锁骨颅骨发育不全相关,发现其polyQ/polyA CC的稳定性控制其聚集、定位和毒性。这些发现表明,与polyQ一样,polyA重复序列在人类遗传疾病中扩增时也会形成CC结构,从而触发蛋白质聚集和毒性。