The Nup107-160 complex and γ-TuRC regulate microtubule polymerization at kinetochores

被引:145
作者
Mishra, Ram Kumar [1 ]
Chakraborty, Papia [2 ]
Arnaoutov, Alexei [1 ]
Fontoura, Beatriz M. A. [2 ]
Dasso, Mary [1 ]
机构
[1] NICHD, Lab Gene Regulat & Dev, NIH, Bethesda, MD 20892 USA
[2] Univ Texas SW Med Ctr Dallas, Dept Cell Biol, Dallas, TX 75390 USA
基金
美国国家卫生研究院;
关键词
XENOPUS EGG EXTRACTS; NUCLEAR-PORE; MAMMALIAN-CELLS; MESSENGER-RNA; SPINDLE; SUBCOMPLEX; TUBULIN; MITOSIS; RAN; ORGANIZATION;
D O I
10.1038/ncb2016
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The metazoan nuclear pore complex (NPC) disassembles during mitosis, and many of its constituents distribute onto spindles and kinetochores, including the Nup107-160 subcomplex(1,2). We have found that Nup107-160 interacts with the gamma-tubulin ring complex (gamma-TuRC), an essential and conserved microtubule nucleator(3,4), and recruits gamma-TuRC to unattached kinetochores. The unattached kinetochores nucleate microtubules in a manner that is regulated by Ran GTPase(5); such microtubules contribute to the formation of kinetochore fibres (k-fibres)(6), microtubule bundles connecting kinetochores to spindle poles. Our data indicate that Nup107-160 and gamma-TuRC act cooperatively to promote spindle assembly through microtubule nucleation at kinetochores: HeLa cells lacking Nup107-160 or gamma-TuRC were profoundly deficient in kinetochore-associated microtubule nucleation. Moreover, co-precipitated Nup107-160-gamma-TuRC complexes nucleated microtubule formation in assays using purified tubulin. Although Ran did not regulate microtubule nucleation by gamma-TuRC alone, Nup107-160-gamma-TuRC complexes required RanGTP for microtubule nucleation. Collectively, our observations show that Nup107-160 promotes spindle assembly through Ran-GTP-regulated nucleation of microtubules by gamma-TuRC at kinetochores, and reveal a relationship between nucleoporins and the microtubule cytoskeleton.
引用
收藏
页码:164 / U154
页数:10
相关论文
共 29 条
[1]   Nup120p: A yeast nucleoporin required for NPC distribution and mRNA transport [J].
Aitchison, JD ;
Blobel, G ;
Rout, MP .
JOURNAL OF CELL BIOLOGY, 1995, 131 (06) :1659-1675
[2]   Crm1 is a mitotic effector of Ran-GTP in somatic cells [J].
Arnaoutov, A ;
Azuma, Y ;
Ribbeck, K ;
Joseph, J ;
Boyarchuk, Y ;
Karpova, T ;
McNally, J ;
Dasso, M .
NATURE CELL BIOLOGY, 2005, 7 (06) :626-U27
[3]   An evolutionarily conserved NPC subcomplex, which redistributes in part to kinetochores in mammalian cells [J].
Belgareh, N ;
Rabut, G ;
Baï, SW ;
van Overbeek, M ;
Beaudouin, J ;
Daigle, N ;
Zatsepina, OV ;
Pasteau, F ;
Labas, V ;
Fromont-Racine, M ;
Ellenberg, J ;
Doye, V .
JOURNAL OF CELL BIOLOGY, 2001, 154 (06) :1147-1160
[4]  
Desai A, 1999, METHOD CELL BIOL, V61, P385
[5]   IDENTIFICATION OF A FAMILY OF HUMAN CENTROMERE PROTEINS USING AUTOIMMUNE SERA FROM PATIENTS WITH SCLERODERMA [J].
EARNSHAW, WC ;
ROTHFIELD, N .
CHROMOSOMA, 1985, 91 (3-4) :313-321
[6]   Augmin: a protein complex required for centrosome-independent microtubule generation within the spindle [J].
Goshima, Gohta ;
Mayer, Mirjam ;
Zhang, Nan ;
Stuurman, Nico ;
Vale, Ronald D. .
JOURNAL OF CELL BIOLOGY, 2008, 181 (03) :421-429
[7]   Genes required for mitotic spindle assembly in Drosophila S2 cells [J].
Goshima, Gohta ;
Wollman, Roy ;
Goodwin, Sarah S. ;
Zhang, Nan ;
Scholey, Jonathan M. ;
Vale, Ronald D. ;
Stuurman, Nico .
SCIENCE, 2007, 316 (5823) :417-421
[8]   Removal of a single pore subcomplex results in vertebrate nuclei devoid of nuclear pores [J].
Harel, A ;
Orjalo, AV ;
Vincent, T ;
Lachish-Zalait, A ;
Vasu, S ;
Shah, S ;
Zimmerman, E ;
Elbaum, M ;
Forbes, DJ .
MOLECULAR CELL, 2003, 11 (04) :853-864
[9]   Self-organization of microtubules into bipolar spindles around artificial chromosomes in Xenopus egg extracts [J].
Heald, R ;
Tournebize, R ;
Blank, T ;
Sandaltzopoulos, R ;
Becker, P ;
Hyman, A ;
Karsenti, E .
NATURE, 1996, 382 (6590) :420-425
[10]   The RanGAP1-RanBP2 complex is essential for microtubule-kinetochore interactions in vivo [J].
Joseph, J ;
Liu, ST ;
Jablonski, SA ;
Yen, TJ ;
Dasso, M .
CURRENT BIOLOGY, 2004, 14 (07) :611-617