Renvoisé Benoît, Colasse Sabrina, Burlet Philippe, Viollet Louis, Meier U Thomas, Lefebvre Suzie
Laboratoire de Biologie Cellulaire des Membranes, Department of Cell Biology, Institut Jacques Monod (IJM), UMR 7592 CNRS/Universités Paris 6 et 7, Paris Cedex 05, France.
Hum Mol Genet. 2009 Apr 1;18(7):1181-9. doi: 10.1093/hmg/ddp009. Epub 2009 Jan 7.
Spinal muscular atrophy (SMA) is a common autosomal recessive neurodegenerative disease caused by reduced survival motor neuron (SMN) levels. The assembly machinery containing SMN is implicated in the biogenesis of the spliceosomal small nuclear ribonucleoproteins (snRNPs). SMN is present in both the cytoplasm and nucleus, where it transiently accumulates in subnuclear domains named Cajal bodies (CBs) and functions in the maturation of snRNPs and small nucleolar (sno)RNPs. The impact of lowering SMN levels on the composition of CBs in SMA cells is still not completely understood. Here, we analyse the CB composition in immortalized and primary fibroblasts from SMA patients. We show that the U snRNA export factors PHAX and chromosome region maintenance 1 and the box C/D snoRNP core protein fibrillarin concentrate in CBs from SMA cells, whereas the box H/ACA core proteins GAR1 and NAP57/dyskerin show reduced CB localization. Remarkably, the functional deficiency in SMA cells is associated with decreased localization of the snoRNP chaperone Nopp140 in CBs that correlates with disease severity. Indeed, RNA interference knockdown experiments in control fibroblasts demonstrate that SMN is required for accumulation of Nopp140 in CBs. Conversely, overexpression of SMN in SMA cells restores the CB localization of Nopp140, whereas SMN mutants found in SMA patients are defective in promoting the association of Nopp140 with CBs. Taken together, we demonstrate that only a subset of CB functions (as indicated by the association of representative factors) are impaired in SMA cells and, importantly, we identify the decrease of Nopp140 localization in CBs as a phenotypic marker for SMA.
脊髓性肌萎缩症(SMA)是一种常见的常染色体隐性神经退行性疾病,由存活运动神经元(SMN)水平降低引起。包含SMN的组装机制与剪接体小核核糖核蛋白(snRNP)的生物合成有关。SMN存在于细胞质和细胞核中,在细胞核中它会短暂积聚在名为 Cajal 体(CB)的亚核区域,并在 snRNP 和小核仁(sno)RNP 的成熟过程中发挥作用。降低SMN水平对SMA细胞中CB组成的影响仍未完全了解。在这里,我们分析了SMA患者永生化和成纤维细胞中的CB组成。我们发现,U snRNA 输出因子 PHAX 和染色体区域维持蛋白 1 以及盒 C/D snoRNP 核心蛋白纤维蛋白原集中在SMA细胞的CB中,而盒 H/ACA 核心蛋白 GAR1 和 NAP57/营养不良症蛋白在CB中的定位减少。值得注意的是,SMA细胞中的功能缺陷与CB中snoRNP伴侣蛋白Nopp140的定位减少有关,这与疾病严重程度相关。事实上,在对照成纤维细胞中进行的RNA干扰敲低实验表明,CB中Nopp140的积累需要SMN。相反,在SMA细胞中过表达SMN可恢复Nopp140在CB中的定位,而在SMA患者中发现的SMN突变体在促进Nopp140与CB的结合方面存在缺陷。综上所述,我们证明在SMA细胞中只有一部分CB功能(由代表性因子的结合表明)受损,重要的是,我们确定CB中Nopp140定位的减少是SMA的一个表型标志物。