Nudel modulates kinetochore association and function of cytoplasmic dynein in M phase.
The microtubule-based motor cytoplasmic dynein/dynactin is a force generator at the kinetochore. It also transports proteins away from kinetochores to spindle poles. Regulation of such diverse functions, however, is poorly understood. We have previously shown that Nudel is critical for dynein-mediated protein transport, whereas mitosin, a kinetochore protein that binds Nudel, is involved in retention of kinetochore dynein/dynactin against microtubule-dependent stripping. Here we demonstrate that Nudel is required for robust localization of dynein/dynactin at the kinetochore. It localizes to kinetochores after nuclear envelope breakdown, depending mostly ( approximately 78%) on mitosin and slightly on dynein/dynactin. Depletion of Nudel by RNA interference (RNAi) or overexpression of its mutant incapable of binding either Lis1 or dynein heavy chain abolishes the kinetochore protein transport and mitotic progression. Similar to mitosin RNAi, Nudel RNAi also leads to increased stripping of kinetochore dynein/dynactin in the presence of microtubules. Taking together, our results suggest a dual role of kinetochore Nudel: it activates dynein-mediated protein transport and, when interacting with both mitosin and dynein, stabilizes kinetochore dynein/dynactin against microtubule-dependent stripping to facilitate the force generation function of the motor.
Liang
Yun
Y
Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Yu
Wei
W
Li
Yan
Y
Yu
Lihou
L
Zhang
Qiangge
Q
Wang
Fubin
F
Yang
Zhenye
Z
Du
Juan
J
Huang
Qiongping
Q
Yao
Xuebiao
X
Zhu
Xueliang
X
D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D002455:Cell Division; D002460:Cell Line; D016764:Cell Polarity; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism*; D000072159:Dynactin Complex; D004398:Dyneins / Q000378:metabolism*; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D008840:Microfilament Proteins / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008954:Models, Biological; D009363:Neoplasm Proteins / Q000378:metabolism; D009685:Nuclear Envelope / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D034622:RNA Interference
D002352:Carrier Proteins; D002868:Chromosomal Proteins, Non-Histone; D000072159:Dynactin Complex; D008840:Microfilament Proteins; D008869:Microtubule-Associated Proteins; C421108:NDEL1 protein, human; D009363:Neoplasm Proteins; C094772:centromere protein F; C076744:cytoplasmic linker protein 170; D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase; C084102:PAFAH1B1 protein, human; D004398:Dyneins