Ganesh Subramaniam, Delgado-Escueta Antonio V, Suzuki Toshimitsu, Francheschetti Silvana, Riggio Concetta, Avanzini Giuiliano, Rabinowicz Adrian, Bohlega Saeed, Bailey Julia, Alonso Maria E, Rasmussen Astrid, Thomson Alfredo E, Ochoa Adriana, Prado Aurelio J, Medina Marco T, Yamakawa Kazuhiro
Laboratory for Neurogenetics, RIKEN Brain Science Institute, Wako-shi, Japan.
Hum Mol Genet. 2002 May 15;11(11):1263-71. doi: 10.1093/hmg/11.11.1263.
Mutations in the EPM2A gene encoding a dual-specificity phosphatase (laforin) cause an autosomal recessive fatal disorder called Lafora's disease (LD) classically described as an adolescent-onset stimulus-sensitive myoclonus, epilepsy and neurologic deterioration. Here we related mutations in EPM2A with phenotypes of 22 patients (14 families) and identified two subsyndromes: (i) classical LD with adolescent-onset stimulus-sensitive grand mal, absence and myoclonic seizures followed by dementia and neurologic deterioration, and associated mainly with mutations in exon 4 (P = 0.0007); (ii) atypical LD with childhood-onset dyslexia and learning disorder followed by epilepsy and neurologic deterioration, and associated mainly with mutations in exon 1 (P = 0.0015). To understand the two subsyndromes better, we investigated the effect of five missense mutations in the carbohydrate-binding domain (CBD-4; coded by exon 1) and three missense mutations in the dual phosphatase domain (DSPD; coded by exons 3 and 4) on laforin's intracellular localization in HeLa cells. Expression of three mutant proteins (T194I, G279S and Y294N) in DSPD formed ubiquitin-positive cytoplasmic aggregates, suggesting that they were folding mutants set for degradation. In contrast, none of the three CBD-4 mutants showed cytoplasmic clumping. However, CBD-4 mutants W32G and R108C targeted both cytoplasm and nucleus, suggesting that laforin had diminished its usual affinity for polysomes. Our data, thus, represent the first report of a novel childhood syndrome for LD. Our results also provide clues for distinct roles for the CBD-4 and DSP domains of laforin in the etiology of two subsyndromes of LD.
编码双特异性磷酸酶(拉福林)的EPM2A基因突变会导致一种常染色体隐性致命疾病,即经典描述为青少年期起病的刺激敏感性肌阵挛、癫痫和神经功能恶化的拉福拉病(LD)。在此,我们将EPM2A基因突变与22例患者(14个家系)的表型相关联,并确定了两种亚综合征:(i)经典型LD,青少年期起病,刺激敏感性全身性癫痫发作、失神发作和肌阵挛发作,随后出现痴呆和神经功能恶化,主要与外显子4中的突变相关(P = 0.0007);(ii)非典型LD,儿童期起病,伴有诵读困难和学习障碍,随后出现癫痫和神经功能恶化,主要与外显子1中的突变相关(P = 0.0015)。为了更好地理解这两种亚综合征,我们研究了碳水化合物结合结构域(CBD - 4;由外显子1编码)中的五个错义突变和双磷酸酶结构域(DSPD;由外显子3和4编码)中的三个错义突变对拉福林在HeLa细胞内定位的影响。DSPD中三种突变蛋白(T194I、G279S和Y294N)的表达形成了泛素阳性的细胞质聚集体,表明它们是准备降解的折叠突变体。相比之下,三种CBD - 4突变体均未显示细胞质聚集。然而,CBD - 4突变体W32G和R108C同时靶向细胞质和细胞核,表明拉福林对多核糖体的亲和力降低。因此,我们的数据代表了关于LD一种新型儿童综合征的首次报道。我们的结果还为拉福林的CBD - 4和DSP结构域在LD两种亚综合征病因中的不同作用提供了线索。