Ohkawara Bisei, Cabrera-Serrano Macarena, Nakata Tomohiko, Milone Margherita, Asai Nobuyuki, Ito Kenyu, Ito Mikako, Masuda Akio, Ito Yasutomo, Engel Andrew G, Ohno Kinji
Division of Neurogenetics, Center for Neurological Diseases and Cancer and.
Hum Mol Genet. 2014 Apr 1;23(7):1856-68. doi: 10.1093/hmg/ddt578. Epub 2013 Nov 13.
Congenital myasthenic syndromes (CMS) are heterogeneous disorders in which the safety margin of neuromuscular transmission is compromised by one or more specific mechanisms. Using Sanger and exome sequencing in a CMS patient, we identified two heteroallelic mutations, p.Glu1233Lys and p.Arg1277His, in LRP4 coding for the postsynaptic low-density lipoprotein receptor-related protein 4. LRP4, expressed on the surface of the postsynaptic membrane of the neuromuscular junction, is a receptor for neurally secreted agrin, and LRP4 bound by agrin activates MuSK. Activated MuSK in concert with Dok-7 stimulates rapsyn to concentrate and anchor AChR on the postsynaptic membrane and interacts with other proteins implicated in the assembly and maintenance of the neuromuscular junction. LRP4 also functions as an inhibitor of Wnt/beta-catenin signaling. The identified mutations in LRP4 are located at the edge of its 3rd beta-propeller domain and decrease binding affinity of LRP4 for both MuSK and agrin. Mutations in the LRP4 3rd beta-propeller domain were previously reported to impair Wnt signaling and cause bone diseases including Cenani-Lenz syndactyly syndrome and sclerosteosis-2. By analyzing naturally occurring and artificially introduced mutations in the LRP4 3rd beta-propeller domain, we show that the edge of the domain regulates the MuSK signaling whereas its central cavity governs Wnt signaling. We conclude that LRP4 is a new CMS disease gene and that the 3rd beta propeller domain of LRP4 mediates the two signaling pathways in a position-specific manner.
先天性肌无力综合征(CMS)是一类异质性疾病,其中神经肌肉传递的安全边际因一种或多种特定机制而受损。在一名CMS患者中使用桑格测序法和外显子组测序,我们在编码突触后低密度脂蛋白受体相关蛋白4(LRP4)的基因中鉴定出两个杂合等位基因突变,即p.Glu1233Lys和p.Arg1277His。LRP4在神经肌肉接头的突触后膜表面表达,是神经分泌的聚集蛋白的受体,与聚集蛋白结合的LRP4会激活肌肉特异性激酶(MuSK)。被激活的MuSK与Dok-7协同作用,刺激rapsyn将乙酰胆碱受体(AChR)聚集并锚定在突触后膜上,并与其他参与神经肌肉接头组装和维持的蛋白质相互作用。LRP4还作为Wnt/β-连环蛋白信号通路的抑制剂发挥作用。所鉴定出的LRP4突变位于其第3个β-螺旋桨结构域的边缘,降低了LRP4对MuSK和聚集蛋白的结合亲和力。先前有报道称,LRP4第3个β-螺旋桨结构域的突变会损害Wnt信号通路,并导致包括塞纳尼-伦茨并指综合征和骨硬化症-2在内的骨骼疾病。通过分析LRP4第3个β-螺旋桨结构域中自然发生和人工引入的突变,我们发现该结构域的边缘调节MuSK信号通路,而其中心腔则控制Wnt信号通路。我们得出结论,LRP4是一种新的CMS疾病基因,并且LRP4的第3个β-螺旋桨结构域以位置特异性方式介导这两种信号通路。