Human CLASP1 is an outer kinetochore component that regulates spindle microtubule dynamics

被引:171
作者
Maiato, H
Fairley, EAL
Rieder, CL
Swedlow, JR
Sunkel, CE
Earnshaw, WC
机构
[1] Univ Edinburgh, Inst Cell & Mol Biol, Chromosome Struct Grp, Wellcome Trust Ctr Cell Biol, Edinburgh EH9 3JR, Midlothian, Scotland
[2] Univ Porto, Inst Biol Mol & Celular, Genet Mol Lab, P-4150180 Oporto, Portugal
[3] Wadsworth Ctr, Lab Cell Regulat, Div Mol Med, Albany, NY 12201 USA
[4] Univ Dundee, Div Gene Regulat & Express, MSI, WTB Complex, Dundee DD1 5EH, Scotland
[5] Univ Porto, Inst Ciencias Biomed Abel Alazar, P-4099003 Oporto, Portugal
基金
英国惠康基金;
关键词
D O I
10.1016/S0092-8674(03)00465-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
One of the most intriguing aspects of mitosis is the ability of kinetochores to hold onto plus ends of microtubules that are actively gaining or losing tubulin subunits. Here, we show that CLASP1, a microtubule-associated protein, localizes preferentially near the plus ends of growing spindle microtubules and is also a component of a kinetochore region that we term the outer corona. A truncated form of CLASP1 lacking the kinetochore binding domain behaves as a dominant negative, leading to the formation of radial arrays of microtubule bundles that are highly resistant to depolymerization. Microinjection of CLASP1-specific antibodies suppresses microtubule dynamics at kinetochores and throughout the spindle, resulting in the formation of monopolar asters with chromosomes buried in the interior. Incubation with microtubule-stabilizing drugs rescues the kinetochore association with microtubule plus ends at the periphery of the asters. Our data suggest that CLASP1 is required at kinetochores for attached microtubules to exhibit normal dynamic behavior.
引用
收藏
页码:891 / 904
页数:14
相关论文
共 56 条
[1]   CENP-E as an essential component of the mitotic checkpoint in vitro [J].
Abrieu, A ;
Kahana, JA ;
Wood, KW ;
Cleveland, DW .
CELL, 2000, 102 (06) :817-826
[2]  
AIZAWA H, 1991, J BIOL CHEM, V266, P9841
[3]   CLASPs are CLIP-115 and-170 associating proteins involved in the regional regulation of microtubule dynamics in motile fibroblasts [J].
Akhmanova, A ;
Hoogenraad, CC ;
Drabek, K ;
Stepanova, T ;
Dortland, B ;
Verkerk, T ;
Vermeulen, W ;
Burgering, BM ;
De Zeeuw, CI ;
Grosveld, F ;
Galjart, N .
CELL, 2001, 104 (06) :923-935
[4]   Comparison of ARM and HEAT protein repeats [J].
Andrade, MA ;
Petosa, C ;
O'Donoghue, SI ;
Müller, CW ;
Bork, P .
JOURNAL OF MOLECULAR BIOLOGY, 2001, 309 (01) :1-18
[5]   Structural transitions at microtubule ends correlate with their dynamic properties in Xenopus egg extracts [J].
Arnal, I ;
Karsenti, E ;
Hyman, AA .
JOURNAL OF CELL BIOLOGY, 2000, 149 (04) :767-774
[6]   FINE STRUCTURE OF KINETOCHORE OF A MAMMALIAN CELL IN VITRO [J].
BRINKLEY, BR ;
STUBBLEFIELD, E .
CHROMOSOMA, 1966, 19 (01) :28-+
[7]   DISTRIBUTIVE SEGREGATION - MOTORS IN THE POLAR WIND [J].
CARPENTER, ATC .
CELL, 1991, 64 (05) :885-890
[8]   Implication of a novel multiprotein Dam1p complex in outer kinetochore function [J].
Cheeseman, IM ;
Brew, C ;
Wolyniak, M ;
Desai, A ;
Anderson, S ;
Muster, N ;
Yates, JR ;
Huffaker, TC ;
Drubin, DG ;
Barnes, G .
JOURNAL OF CELL BIOLOGY, 2001, 155 (07) :1137-1145
[9]   Mitotic spindle integrity and kinetochore function linked by the Duo1p/Dam1p complex [J].
Cheeseman, IM ;
Enquist-Newman, M ;
Müller-Reichert, T ;
Drubin, DG ;
Barnes, G .
JOURNAL OF CELL BIOLOGY, 2001, 152 (01) :197-212
[10]   CENP-B - A MAJOR HUMAN CENTROMERE PROTEIN LOCATED BENEATH THE KINETOCHORE [J].
COOKE, CA ;
BERNAT, RL ;
EARNSHAW, WC .
JOURNAL OF CELL BIOLOGY, 1990, 110 (05) :1475-1488