Biochemical characterization and cellular effects of CADASIL mutants of NOTCH3.
Biochemical characterization and cellular effects of CADASIL mutants of NOTCH3.
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DOI：
10.1371/journal.pone.0044964
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Wang MM
中科院分区：
文献类型：
--
作者：
Meng H;Zhang X;Yu G;Lee SJ;Chen YE;Prudovsky I;Wang MM
常染色体显性遗传性脑动脉病伴皮质下白质脑病（CADASIL）是显性遗传性卒中的最佳病因，由导致NOTCH 3蛋白蓄积和选择性动脉平滑肌变性的NOTCH 3突变引起。先前的研究表明，NOTCH 3蛋白形成多聚体。在这里，我们研究NOTCH 3和其他血管Notch亚型之间的蛋白质相互作用，并表征升高的NOTCH 3对平滑肌基因调控的影响。我们证明了NOTCH 3与NOTCH 1、NOTCH 3和NOTCH 4形成异二聚体。R90 C和C49 Y突变体N 0 TCH 3形成的复合物比野生型N 0 TCH 3复合物对去污剂更具抗性。使用定量NOTCH 3-荧光素酶清除测定，我们发现突变体NOTCH 3清除的显著抑制。在NOTCH功能的共培养试验中，过表达的野生型和突变型NOTCH 3显著抑制了NOTCH调节的平滑肌转录物，并有力地损害了三个独立的平滑肌启动子的活性。应用于细胞培养物的野生型和R90 C重组N 0 TCH 3蛋白也阻断典型Notch功能。我们得出结论，NOTCH 3的CADASIL突变体与NOTCH 1、3和4复合，减缓NOTCH 3清除，并且过表达的野生型和突变型NOTCH 3蛋白干扰平滑肌细胞中关键的NOTCH介导的功能。
Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) is the best understood cause of dominantly inherited stroke and results from NOTCH3 mutations that lead to NOTCH3 protein accumulation and selective arterial smooth muscle degeneration. Previous studies show that NOTCH3 protein forms multimers. Here, we investigate protein interactions between NOTCH3 and other vascular Notch isoforms and characterize the effects of elevated NOTCH3 on smooth muscle gene regulation. We demonstrate that NOTCH3 forms heterodimers with NOTCH1, NOTCH3, and NOTCH4. R90C and C49Y mutant NOTCH3 form complexes which are more resistant to detergents than wild type NOTCH3 complexes. Using quantitative NOTCH3-luciferase clearance assays, we found significant inhibition of mutant NOTCH3 clearance. In coculture assays of NOTCH function, overexpressed wild type and mutant NOTCH3 significantly repressed NOTCH-regulated smooth muscle transcripts and potently impaired the activity of three independent smooth muscle promoters. Wildtype and R90C recombinant NOTCH3 proteins applied to cell cultures also blocked canonical Notch fuction. We conclude that CADASIL mutants of NOTCH3 complex with NOTCH1, 3, and 4, slow NOTCH3 clearance, and that overexpressed wild type and mutant NOTCH3 protein interfere with key NOTCH-mediated functions in smooth muscle cells.
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影响因子：
9.9
作者：
CHABRIAT, H;TOURNIERLASSERVE, E;BOUSSER, MG
通讯作者：
BOUSSER, MG
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：
64.8
作者：
JARRIAULT, S;BROU, C;ISRAEL, A
通讯作者：
ISRAEL, A
影响因子：
11.2
作者：
Dichgans, M;Mayer, M;Gasser, T
通讯作者：
Gasser, T
影响因子：
10.5
作者：
Domenga, V;Fardoux, P;Joutel, A
通讯作者：
Joutel, A