Transendocytosis is impaired in CADASIL-mutant NOTCH3
Transendocytosis is impaired in CADASIL-mutant NOTCH3
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CADASIL 突变型 NOTCH3 的转内吞作用受损
DOI：
10.1016/j.expneurol.2011.10.020
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发表时间：
2012
影响因子：
5.3
通讯作者：
Mizuno T
中科院分区：
文献类型：
--
作者：
Watanabe-Hosomi A;Watanabe Y;Tanaka M;Nakagawa M;Mizuno T
人类NOTCH 3基因突变导致伴有皮质下梗死和白质脑病的常染色体显性遗传性脑动脉病（CADASIL），但CADASIL的发病机制仍不清楚。最近，Notch外胚层结构域内吞到配体表达细胞中，称为转内吞作用，已被认为是Notch激活的关键。我们假设突变的NOTCH3蛋白，特别是NOTCH3的外胚层结构域（N3ECD），可能由于受损的转内吞作用而难以在细胞表面降解。我们建立了一个共培养系统，其中稳定表达一个拷贝的四环素调节的NOTCH3的HEK293细胞与表达NOTCH3配体Jagged1（Jag1）的HEK293细胞一起培养。我们获得了三个主要结果：第一，当NOTCH3细胞与Jag1表达细胞共培养时，细胞表面的C185 R突变体N3ECD比野生型N3ECD降解得更慢。第二，野生型和突变型NOTCH 3表达细胞在与配体表达细胞共培养时均增加了HES 1表达。第三，在Jag1表达细胞中观察到含有N3ECD的囊泡。囊泡的突变N3ECD内的Jag1表达细胞的数量显着少于野生型N3ECD的情况下。这些结果表明，由于转内吞功能受损，突变型N3ECD在细胞表面的降解过程受到干扰。血管平滑肌细胞表面的这种紊乱可能有助于CADASIL的发病机制。
Mutations in the human NOTCH3 gene cause cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), but the pathogenesis of CADASIL has remained unclear. Recently, endocytosis of the Notch ectodermal domain into ligand-expressing cells, called transendocytosis, has come to be considered critical for Notch activation. We hypothesized that the mutant NOTCH3 protein, particularly the ectodermal domain of NOTCH3 (N3ECD), may be refractory to degradation on the cell surface due to impaired transendocytosis. We established a co-culture system in which HEK293 cells stably expressing one copy of tetracycline-regulated NOTCH3 were cultured with NOTCH3 ligand Jagged1 (Jag1)-expressing HEK293 cells. We obtained three main results: first, the C185R mutant N3ECD on the cell surface was degraded significantly more slowly than the wild N3ECD when NOTCH3 cells were co-cultured with Jag1-expressing cells. Second, both the wild-type and mutant NOTCH3-expressing cells increased HES1 expression on co-culture with ligand-expressing cells. Third, vesicles containing N3ECD were observed in Jag1-expressing cells. Vesicles of mutant N3ECD within the Jag1-expressing cells were significantly less in number than in the case of wild-type N3ECD. These results indicated that the process of degradation of mutant N3ECD on the cell surface is disturbed due to impairment of transendocytosis. Such disturbance on the surface of vascular smooth muscle cells may contribute to the pathogenesis of CADASIL.
DOI：
10.1073/pnas.252624099
发表时间：
2002-12-24
影响因子：
11.1
作者：
Karlström, H;Beatus, P;Lundkvist, J
通讯作者：
Lundkvist, J
影响因子：
10.5
作者：
Domenga, V;Fardoux, P;Joutel, A
通讯作者：
Joutel, A
细胞给予和细胞带走：通过配体和受体的内吞运输激活Notch通路的概述。
影响因子：
2.5
作者：
Pratt,EmilyB;Wentzell,JillS;Maxson,JuliaE;Courter,Lauren;Hazelett,Dennis;Christian,JanL
通讯作者：
Christian,JanL