Kinetochore Microtubule Dynamics and Attachment Stability Are Regulated by Hec1

DeLuca, Jennifer G.; Gall, Walter; Ciferri, Claudio; Cimini, Daniela; Musacchio, Andrea; Salmon, E. D.

doi:10.1016/j.cell.2006.09.047

Cited by 698 publications

(863 citation statements)

References 55 publications

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Section: Control Of Kinetochore Mt Dynamicsmentioning

confidence: 67%

“…Our data are consistent with the recent findings of Wordeman et al (2007), who propose an attractive idea that the role of MCAK is to loosen up MT ends to facilitate directional coordination of chromosome motility and MT release. Another interesting observation is that although it has been clearly established that MCAK is a substrate of Aurora B kinase (Andrews et al, 2004;Lan et al, 2004;Sampath et al, 2004;Zhang et al, 2007), the magnitude of the effects of MCAK inhibition versus Aurora B inhibition on the t 1/2 value of the K-fiber strongly supports the idea that Aurora B causes release of aberrant MT attachments primarily through a mechanism that does not rely solely on MCAK DeLuca et al, 2006;Wordeman et al, 2007).Overall our work supports a model where MCAK differentially affects the MT subclasses which coexist during mitosis, with specific effects on K-fibers and astral MTs. This is consistent with the complex localization of MCAK in the spindle and the highly complicated phosphorregulatory schemes that regulate its localization and activity at centromeres and at spindle poles (Andrews et al, 2004;Gorbsky, 2004;Lan et al, 2004;Ohi et al, 2004;Knowlton et al, 2006;Zhang et al, 2007Zhang et al, , 2008.…”

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confidence: 70%

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MCAK and Paclitaxel Have Differential Effects on Spindle Microtubule Organization and Dynamics

Rizk

Bohannon

Wetzel

et al. 2009

MBoC

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Within the mitotic spindle, there are multiple populations of microtubules with different turnover dynamics, but how these different dynamics are maintained is not fully understood. MCAK is a member of the kinesin-13 family of microtubule-destabilizing enzymes that is required for proper establishment and maintenance of the spindle. Using quantitative immunofluorescence and fluorescence recovery after photobleaching, we compared the differences in spindle organization caused by global suppression of microtubule dynamics, by treating cells with low levels of paclitaxel, versus specific perturbation of spindle microtubule subsets by MCAK inhibition. Paclitaxel treatment caused a disruption in spindle microtubule organization marked by a significant increase in microtubules near the poles and a reduction in K-fiber fluorescence intensity. This was correlated with a faster t 1/2 of both spindle and K-fiber microtubules. In contrast, MCAK inhibition caused a dramatic reorganization of spindle microtubules with a significant increase in astral microtubules and reduction in K-fiber fluorescence intensity, which correlated with a slower t 1/2 of K-fibers but no change in the t 1/2 of spindle microtubules. Our data support the model that MCAK perturbs spindle organization by acting preferentially on a subset of microtubules, and they support the overall hypothesis that microtubule dynamics is differentially regulated in the spindle. INTRODUCTIONThe cytoskeleton must undergo dramatic reorganization as cells transition from interphase to mitosis to assemble the mitotic spindle. The spindle is a macromolecular structure composed of a dynamic array of microtubules (MTs) and their associated proteins that is necessary for proper chromosome segregation (Compton, 2000;Walczak and Heald, 2008). At the onset of mitosis, there is an increase in MT turnover that allows for breakdown of the interphase array and assembly of the mitotic spindle (Saxton et al., 1984;. This change is correlated with an increased catastrophe frequency (transition from growth to shrinkage) and a decreased rescue frequency (transition from shrinkage to growth) (Belmont et al., 1990;Gliksman et al., 1992;Verde et al., 1992;Rusan et al., 2001). The changes in MT dynamics are also correlated with a change in MT polymer levels during mitosis (Zhai et al., 1996). At nuclear envelope breakdown, there is a dramatic decrease in MT polymer levels, which may allow the cell to rapidly assemble a mitotic spindle because the released tubulin dimers can polymerize into new spindle MTs. As the cell progresses from mitosis to the next interphase, there is no change in the levels of MT polymer (Zhai et al., 1996), suggesting that MT polymer gets reorganized to reform the interphase MT cytoskeleton, without a significant change in the dynamics of the MTs. These studies highlight the need for the cell to possess a mechanism to temporally regulate MT dynamics.In addition to the temporal changes in dynamics throughout the cell cycle, the dynamics of MTs within mitosis are also...

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Section: Control Of Kinetochore Mt Dynamicsmentioning

confidence: 67%

mentioning

confidence: 70%

MCAK and Paclitaxel Have Differential Effects on Spindle Microtubule Organization and Dynamics

Rizk

Bohannon

Wetzel

et al. 2009

MBoC

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“…However, NEK2A is not required for assembly of Hec1 to kinetochore (Figures 2A-C), suggesting that other regulatory mechanisms exist to govern the assembly of Ndc80 complex. Two recent elegant studies illustrated that mitotic kinase Aurora B also regulates the globular domain of Hec1 (Cheeseman et al, 2006;DeLuca et al, 2006), which reduces the affinity of the Ndc80 complex for microtubules in vitro and perhaps corrects improper kinetochoremicrotubule connections in vivo. Therefore, it would be of great interest to elucidate the respective contribution of Aurora B and NEK2A in orchestrating kinetochore-microtubule attachment in real-time chromosome movements.…”

Section: Discussionmentioning

confidence: 99%

“…Another process to prevent errors is to stabilize and destabilize the kinetochore-microtubule connection (Cleveland et al, 2003). Mounting evidence demonstrates that the Ndc80 complex, comprised of Hec1 (highly expressed in cancer), Nuf2, Spc24 and Spc25, constitutes an important kinetochore-microtubule connection in vivo and in vitro (Cheeseman et al, 2006;DeLuca et al, 2006). Hec1 was initially identified as an Rb-binding protein and subsequently demonstrated as an evolutionarily conserved kinetochore protein (Durfee et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

The mitotic checkpoint kinase NEK2A regulates kinetochore microtubule attachment stability

Cai

Yao

et al. 2008

Oncogene

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Loss or gain of whole chromosome, the form of chromosome instability commonly associated with cancers is thought to arise from aberrant chromosome segregation during cell division. Chromosome segregation in mitosis is orchestrated by the interaction of kinetochores with spindle microtubules. Our studies show that NEK2A is a kinetochore-associated protein kinase essential for faithful chromosome segregation. However, it was unclear how NEK2A ensures accurate chromosome segregation in mitosis. Here we show that NEK2A-mediated Hec1 (highly expressed in cancer) phosphorylation is essential for faithful kinetochore microtubule attachments in mitosis. Using phospho-specific antibody, our studies show that NEK2A phosphorylates Hec1 at Ser165 during mitosis. Although such phosphorylation is not required for assembly of Hec1 to the kinetochore, expression of non-phosphorylatable mutant Hec1 S165 perturbed chromosome congression and resulted in a dramatic increase in microtubule attachment errors, including syntelic and monotelic attachments. Our in vitro reconstitution experiment demonstrated that Hec1 binds to microtubule in low affinity and phosphorylation by NEK2A, which prevents aberrant kinetochore-microtubule connections in vivo, increases the affinity of the Ndc80 complex for microtubules in vitro. Thus, our studies illustrate a novel regulatory mechanism in which NEK2A kinase operates a faithful chromosome attachment to spindle microtubule, which prevents chromosome instability during cell division.

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“…The overall conclusion of these studies was that the core of eukaryotic kinetochores is composed of two large conserved protein networks: on one side the KMN network, which consists of Knl-1, the hetero-tetrameric MIND/Mis12 subcomplex, and the hetero-tetrameric-NDC80 subcomplex, and on the other side the CCAN network (Constitutive Centromere Associated Network, which is also called CENP-A NAC/CAD or CENP-H/I complex), which consists of 15 subunits. Soon after the NDC80 subcomplex was recognised as the key microtubule attachment site at kinetochores, which is essential for the binding to spindle microtubules in all eukaryotes DeLuca et al, 2006).…”

Section: Kinetochores a Highly Conserved Structure Only At Second Lookmentioning

confidence: 99%

Keeping kinetochores on track

Meraldi

2012

European Journal of Cell Biology

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a b s t r a c tThe multiple functions of kinetochores are reflected in their complex composition, with over a hundred different proteins, which self-associate in several functional subcomplexes. Most of these kinetochore proteins were identified over the last 10-12 years using a combination of genetic, cell biological, biochemical, and bioinformatic approaches in various model organisms. The key challenge since then has been to determine the structural architecture of kinetochores, define the functions of its different subcomponents, and understand its regulation, both in response to the rapid changes in microtubule dynamics or to sense erroneous attachments for spindle checkpoint signalling. Here, we present some of the key advances obtained in the last six years on the biology of kinetochores, both through our work and through the work of many other groups studying this exciting structure.

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Kinetochore Microtubule Dynamics and Attachment Stability Are Regulated by Hec1

Cited by 698 publications

References 55 publications

MCAK and Paclitaxel Have Differential Effects on Spindle Microtubule Organization and Dynamics

MCAK and Paclitaxel Have Differential Effects on Spindle Microtubule Organization and Dynamics

The mitotic checkpoint kinase NEK2A regulates kinetochore microtubule attachment stability

Keeping kinetochores on track

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