De novo mutations in KIF1A-associated neuronal disorder (KAND) dominant-negatively inhibit motor activity and axonal transport of synaptic vesicle precursors.
De novo mutations in KIF1A-associated neuronal disorder (KAND) dominant-negatively inhibit motor activity and axonal transport of synaptic vesicle precursors.
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DOI：
10.1073/pnas.2113795119
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发表时间：
2022-08-09
影响因子：
11.1
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中科院分区：
文献类型：
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KIF 1A在轴突中转运突触囊泡前体。最近的研究已经在先天性神经病患者中发现了许多KIF 1A突变;然而，发病机制的分子机制在很大程度上仍然是难以捉摸的。本研究建立了秀丽隐杆线虫KIF 1A相关神经元疾病（KAND）的功能丧失模型，以分析体内疾病的分子和细胞生物学。使用该疾病模型进行的遗传筛选鉴定了一种抑制突变，该突变恢复了突变的KIF 1A的运动活性。本研究还建立了体外单分子测定，以定量分析KAND突变对由KIF 1A突变体和野生型拷贝组成的异二聚体马达的影响。我们的研究结果为未来的基因和药物筛选提供了基础，以确定新的KAND疗法。KIF 1A是一种驱动蛋白超家族运动蛋白，在轴突中转运突触囊泡前体。货物结合刺激KIF 1A分子的二聚化以诱导沿着微管的进行性运动。人类Kif 1a的突变导致一组称为KIF 1A相关神经元疾病（KAND）的神经退行性疾病。KAND突变大多是从头和常染色体显性的;然而，尚不清楚野生型KIF 1A马达的功能是否被突变的KIF 1A的异源二聚化抑制。在这里，我们使用CRISPR-Cas9技术建立了KAND的秀丽隐杆线虫模型，并分析了人类KIF 1A突变对轴突运输的影响。在我们的C。elegans模型中，杂合子和纯合子均表现出轴突运输减少。使用疾病模型的抑制剂筛选鉴定了恢复突变的人KIF 1A的运动活性的突变。此外，我们开发了体外试验来分析由野生型和突变型KIF 1A组成的异二聚体马达的运动性。我们发现，突变KIF 1A显着损害异二聚体马达的运动。我们的数据提供了深入了解的分子机制的显性性质的从头KAND突变。
KIF1A transports synaptic vesicle precursors in axons. Recent studies have identified many KIF1A mutations in congenital neuropathy patients; however, the molecular mechanism of pathogenesis remains largely elusive. This study established loss-of-function models for KIF1A-associated neuronal disorder (KAND) in Caenorhabditis elegans to analyze the molecular and cell biology of the disease in vivo. Genetic screening using the disease model identified a suppressor mutation that recovers the motor activity of mutated KIF1A. This study also established in vitro single-molecule assays to quantitatively analyze the effect of KAND mutations on heterodimeric motors composed of mutant and wild-type copies of KIF1A. Our findings provide a foundation for future genetic and drug screening in the effort to identify novel KAND therapies. KIF1A is a kinesin superfamily motor protein that transports synaptic vesicle precursors in axons. Cargo binding stimulates the dimerization of KIF1A molecules to induce processive movement along microtubules. Mutations in human Kif1a lead to a group of neurodegenerative diseases called KIF1A-associated neuronal disorder (KAND). KAND mutations are mostly de novo and autosomal dominant; however, it is unknown if the function of wild-type KIF1A motors is inhibited by heterodimerization with mutated KIF1A. Here, we have established Caenorhabditis elegans models for KAND using CRISPR-Cas9 technology and analyzed the effects of human KIF1A mutation on axonal transport. In our C. elegans models, both heterozygotes and homozygotes exhibited reduced axonal transport. Suppressor screening using the disease model identified a mutation that recovers the motor activity of mutated human KIF1A. In addition, we developed in vitro assays to analyze the motility of heterodimeric motors composed of wild-type and mutant KIF1A. We find that mutant KIF1A significantly impaired the motility of heterodimeric motors. Our data provide insight into the molecular mechanism underlying the dominant nature of de novo KAND mutations.
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影响因子：
16.2
作者：
Klassen, Matthew P.;Wu, Ye E.;Maeder, Celine I.;Nakae, Isei;Cueva, Juan G.;Lehrman, Emily K.;Tada, Minoru;Gengyo-Ando, Keiko;Wang, George J.;Goodman, Miriam;Mitani, Shohei;Kontani, Kenji;Katada, Toshiaki;Shen, Kang
通讯作者：
Shen, Kang
影响因子：
7.8
作者：
HEUSER, JE;REESE, TS
通讯作者：
REESE, TS
DOI：
10.1083/jcb.200908075
发表时间：
2009-12-28
期刊：
The Journal of cell biology
影响因子：
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作者：
Ally S;Larson AG;Barlan K;Rice SE;Gelfand VI
通讯作者：
Gelfand VI
影响因子：
11.4
作者：
Kikkawa, Masahide;Hirokawa, Nobutaka
通讯作者：
Hirokawa, Nobutaka
DOI：
10.1038/nrm2804
发表时间：
2009-12
期刊：
Nature reviews. Molecular cell biology
影响因子：
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