TY - JOUR
T1 - Human CLASP1 is an outer kinetochore component that regulates spindle microtubule dynamics
AU - Maiato, Helder
AU - Fairley, Elizabeth A.L.
AU - Rieder, Conly L.
AU - Swedlow, Jason R.
AU - Sunkel, Claudio E.
AU - Earnshaw, William C.
N1 - Funding Information:
We thank Inke Nathke, Tim Yen, and Anna Akhmanova for the gift of antibodies; Takahiro Nagase and Shoichiro Tsukita for providing KIAA0622 and EB1-GFP cDNAs, respectively; Stefanie Kandels-Lewis and Eric Karsenti for the gift of the HeLa GFP-α-tubulin stable cell line; John Findlay, Richard Cole, and Polla Hergert for help with electron microscopy; and Jorge Vieira for advice on statistical analysis. This work was supported by grants from Fundação para a Ciência e a Tecnologia of Portugal and the TMR Program of the EU to C.E.S., a studentship from the Gulbenkian PhD Programme in Biology and Medicine and FCT to H.M., NIH grant GM 40198 to C.L.R., and grants from The Wellcome Trust to J.R.S. and W.C.E. J.R.S. and W.C.E. are Senior and Principal Research Fellows of The Wellcome Trust, respectively.
PY - 2003/6/27
Y1 - 2003/6/27
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0038385019&partnerID=8YFLogxK
U2 - 10.1016/S0092-8674(03)00465-3
DO - 10.1016/S0092-8674(03)00465-3
M3 - Article
C2 - 12837247
AN - SCOPUS:0038385019
SN - 0092-8674
VL - 113
SP - 891
EP - 904
JO - Cell
JF - Cell
IS - 7
ER -