Nesprin-1α-Dependent Microtubule Nucleation from the Nuclear Envelope via Akap450 Is Necessary for Nuclear Positioning in Muscle Cells.
Nesprin-1α-Dependent Microtubule Nucleation from the Nuclear Envelope via Akap450 Is Necessary for Nuclear Positioning in Muscle Cells.
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DOI：
10.1016/j.cub.2017.08.031
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发表时间：
2017-10-09
期刊：
影响因子：
--
通讯作者：
Gomes ER
中科院分区：
文献类型：
--
作者：
Gimpel P;Lee YL;Sobota RM;Calvi A;Koullourou V;Patel R;Mamchaoui K;Nédélec F;Shackleton S;Schmoranzer J;Burke B;Cadot B;Gomes ER
细胞核是肌细胞中主要的微管组织中心（MTOC），这是由于中心体蛋白的积累和微管（MT）在核膜（NE）上的成核活性。包括Pericentrin、Pcm 1和γ-微管蛋白在内的中心体蛋白的重新定位依赖于Nesprin-1，Nesprin-1是一种通过其N-末端区域将细胞核连接到细胞骨架的外核膜（ONM）蛋白。Nesprin还参与向NE募集驱动蛋白，并在骨骼肌细胞的核定位中发挥作用。然而，MT成核的NE在核定位的功能尚未建立。使用邻近依赖性生物素鉴定（BioID）方法，我们发现了几种中心体蛋白，包括Akap 450、Pcm 1和Pericentrin，其与Nesprin-1α的结合在分化的肌管中增加。我们发现Nesprin-1α不依赖于驱动蛋白而将Akap 450募集到NE，并且Akap 450而不是其他中心体蛋白是NE的MT成核所需的。此外，我们证明，这种机制是破坏先天性肌营养不良症患者肌管携带无义突变的SYNE 1基因（23560 G>T）编码Nesprin-1。最后，使用计算机模拟和细胞培养系统，我们提供了证据的作用，MT成核NE在肌管核扩散。因此，我们的数据揭示了Nesprin-1α/Nesprin-1通过募集Akap 450介导的MT成核活性到NE中在核定位中的新功能。Nesprin-1α的BioID鉴定了肌管核膜上的中心体蛋白Nesprin-1α-含有LINC的复合物将Akap 450募集到肌管核膜上Akap 450是核膜上微管成核所必需的核膜上微管成核参与核定位Gimpel et al.发现包含Nesprin-1α和Sun 1/2的LINC复合物是募集中心体蛋白和Akap 450介导的微管成核活性到肌管核膜所必需的。在体外和计算机模拟中，细胞核中微管成核的缺失影响了肌细胞的核定位。
The nucleus is the main microtubule-organizing center (MTOC) in muscle cells due to the accumulation of centrosomal proteins and microtubule (MT) nucleation activity at the nuclear envelope (NE). The relocalization of centrosomal proteins, including Pericentrin, Pcm1, and γ-tubulin, depends on Nesprin-1, an outer nuclear membrane (ONM) protein that connects the nucleus to the cytoskeleton via its N-terminal region. Nesprins are also involved in the recruitment of kinesin to the NE and play a role in nuclear positioning in skeletal muscle cells. However, a function for MT nucleation from the NE in nuclear positioning has not been established. Using the proximity-dependent biotin identification (BioID) method, we found several centrosomal proteins, including Akap450, Pcm1, and Pericentrin, whose association with Nesprin-1α is increased in differentiated myotubes. We show that Nesprin-1α recruits Akap450 to the NE independently of kinesin and that Akap450, but not other centrosomal proteins, is required for MT nucleation from the NE. Furthermore, we demonstrate that this mechanism is disrupted in congenital muscular dystrophy patient myotubes carrying a nonsense mutation within the SYNE1 gene (23560 G>T) encoding Nesprin-1. Finally, using computer simulation and cell culture systems, we provide evidence for a role of MT nucleation from the NE on nuclear spreading in myotubes. Our data thus reveal a novel function for Nesprin-1α/Nesprin-1 in nuclear positioning through recruitment of Akap450-mediated MT nucleation activity to the NE. BioID of Nesprin-1α identifies centrosomal proteins at myotube nuclear envelope Nesprin-1α-containing LINC complexes recruit Akap450 to myotube nuclear envelope Akap450 is required for microtubule nucleation at the nuclear envelope Microtubule nucleation at the nuclear envelope is involved in nuclear positioning Gimpel et al. find that LINC complexes comprising Nesprin-1α and Sun1/2 are required for the recruitment of centrosomal proteins and Akap450-mediated microtubule nucleation activity to the myotube nuclear envelope. Absence of microtubule nucleation from the nucleus affects nuclear positioning in muscle cells in vitro and in computer simulations.
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DOI：
10.1083/jcb.200301147
发表时间：
2003-06-23
期刊：
The Journal of cell biology
影响因子：
--
作者：
Janson ME;de Dood ME;Dogterom M
通讯作者：
Dogterom M
影响因子：
7.7
作者：
Chojnowski A;Ong PF;Wong ES;Lim JS;Mutalif RA;Navasankari R;Dutta B;Yang H;Liow YY;Sze SK;Boudier T;Wright GD;Colman A;Burke B;Stewart CL;Dreesen O
通讯作者：
Dreesen O
影响因子：
2.8
作者：
Lehmann, Martin;Lichtner, Gregor;Schmoranzer, Jan
通讯作者：
Schmoranzer, Jan
影响因子：
2.7
作者：
Lampe, Andre;Haucke, Volker;Schmoranzer, Jan
通讯作者：
Schmoranzer, Jan
影响因子：
3.3
作者：
Fong, Ka-Wing;Choi, Yuk-Kwan;Qi, Robert Z.
通讯作者：
Qi, Robert Z.