Functional assays for the assessment of the pathogenicity of variants of GOSR2, an ER-to-Golgi SNARE involved in progressive myoclonus epilepsies.
Functional assays for the assessment of the pathogenicity of variants of GOSR2, an ER-to-Golgi SNARE involved in progressive myoclonus epilepsies.
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DOI：
10.1242/dmm.029132
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发表时间：
2017-12-19
影响因子：
4.3
通讯作者：
Schwake M
中科院分区：
文献类型：
--
作者：
Völker JM;Dergai M;Abriata LA;Mingard Y;Ysselstein D;Krainc D;Dal Peraro M;Fischer von Mollard G;Fasshauer D;Koliwer J;Schwake M
进行性肌阵挛癫痫（PMEs）是一种遗传性疾病，其特征是肌阵挛、全身强直阵挛发作和共济失调。与PME相关的基因之一是ER-高尔基体Qb-SNARE GOSR 2，它与syntaxin-5，Bet 1和Sec 22 b形成SNARE复合物。大多数PME患者是p.Gly144Trp突变的纯合子，并出现相似的临床表现。最近，一个病人谁是复合杂合子的p.Gly144Trp和以前看不见的p.Lys164del突变被确定。由于该患者表现为较轻的疾病表型，我们假设p.Lys164del突变可能比p.Gly144Trp突变不那么严重。为了表征p.Gly144Trp和p.Lys164del突变的影响，这两个突变都存在于GOSR 2的SNARE基序中，我们检查了酵母直向同源物Bos 1中的相应突变。在Bos 1中表达正向突变体的酵母表现出受损的生长，表明部分功能丧失，这对于Bos 1 p.Gly176Trp突变更为严重。使用各向异性和凝胶过滤，我们报告，Bos 1 p.Gly176Trp和p.Arg196del能够形成复合物，但具有部分降低的活性。分子动力学（MD）模拟表明，疏水性的核心，触发陷阱复合物的形成，是妥协，由于在GOSR 2和Bos 1的甘氨酸到色氨酸取代。相反，缺失残基p.Lys164（或Bos 1中的p.Arg196del）会干扰GOSR 2和syntaxin-5之间氢键的形成。尽管有这些扰动，所有的SNARE复合体在较长的模拟过程中保持完整。因此，我们的数据表明，在复合杂合子PME中，病程较轻是由于SNARE功能受损较轻。总结：Qb-SNARE GOSR 2突变导致进行性肌阵挛癫痫。作者报告了两种突变对SNARE功能的影响，以研究它们与疾病进展和严重程度的相关性。
Progressive myoclonus epilepsies (PMEs) are inherited disorders characterized by myoclonus, generalized tonic-clonic seizures, and ataxia. One of the genes that is associated with PME is the ER-to-Golgi Qb-SNARE GOSR2, which forms a SNARE complex with syntaxin-5, Bet1 and Sec22b. Most PME patients are homozygous for a p.Gly144Trp mutation and develop similar clinical presentations. Recently, a patient who was compound heterozygous for p.Gly144Trp and a previously unseen p.Lys164del mutation was identified. Because this patient presented with a milder disease phenotype, we hypothesized that the p.Lys164del mutation may be less severe compared to p.Gly144Trp. To characterize the effect of the p.Gly144Trp and p.Lys164del mutations, both of which are present in the SNARE motif of GOSR2, we examined the corresponding mutations in the yeast ortholog Bos1. Yeasts expressing the orthologous mutants in Bos1 showed impaired growth, suggesting a partial loss of function, which was more severe for the Bos1 p.Gly176Trp mutation. Using anisotropy and gel filtration, we report that Bos1 p.Gly176Trp and p.Arg196del are capable of complex formation, but with partly reduced activity. Molecular dynamics (MD) simulations showed that the hydrophobic core, which triggers SNARE complex formation, is compromised due to the glycine-to-tryptophan substitution in both GOSR2 and Bos1. In contrast, the deletion of residue p.Lys164 (or p.Arg196del in Bos1) interferes with the formation of hydrogen bonds between GOSR2 and syntaxin-5. Despite these perturbations, all SNARE complexes stayed intact during longer simulations. Thus, our data suggest that the milder course of disease in compound heterozygous PME is due to less severe impairment of the SNARE function. Summary: Mutations in the Qb-SNARE GOSR2 cause progressive myoclonus epilepsies. The authors report the effect of two mutations on SNARE function to investigate their correlation with progression and severity of disease.
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DOI：
10.1083/jcb.141.7.1489
发表时间：
1998-06-29
期刊：
The Journal of cell biology
影响因子：
--
作者：
通讯作者：
--
影响因子：
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作者：
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影响因子：
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作者：
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作者：
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通讯作者：
D'Adamo, Patrizia
影响因子：
64.5
作者：
Hay, JC;Chao, DS;Scheller, RH
通讯作者：
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