Dynein/dynactin regulate metaphase spindle length by targeting depolymerizing activities to spindle poles

Gaetz, Jedidiah; Kapoor, Tarun M.

doi:10.1083/jcb.200404015

Cited by 184 publications

(207 citation statements)

References 31 publications

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“…Because of technical limitations of the gene knockdown approach, we cannot exclude the possibility that other kinesins and/or dynein might also be involved in mitosis and cytokinesis. Indeed, dynein has previously been implicated in spindle formation and mitosis (Echeverri et al, 1996;Goshima and Vale, 2003;Gaetz and Kapoor, 2004). However, our study and others (Kittler et al, 2004) indicate that depletion of dynein by siRNA does not cause any mitotic or cytokinetic abnormalities.…”

Section: Discussioncontrasting

confidence: 53%

Functional Analysis of Human Microtubule-based Motor Proteins, the Kinesins and Dyneins, in Mitosis/Cytokinesis Using RNA Interference

Zhu

Zhao

Bibikova

et al. 2005

MBoC

374

344

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Microtubule (MT)-based motor proteins, kinesins and dyneins, play important roles in multiple cellular processes including cell division. In this study, we describe the generation and use of an Escherichia coli RNase III-prepared human kinesin/dynein esiRNA library to systematically analyze the functions of all human kinesin/dynein MT motor proteins. Our results indicate that at least 12 kinesins are involved in mitosis and cytokinesis. Eg5 (a member of the kinesin-5 family), Kif2A (a member of the kinesin-13 family), and KifC1 (a member of the kinesin-14 family) are crucial for spindle formation; KifC1, MCAK (a member of the kinesin-13 family), CENP-E (a member of the kinesin-7 family), Kif14 (a member of the kinesin-3 family), Kif18 (a member of the kinesin-8 family), and Kid (a member of the kinesin-10 family) are required for chromosome congression and alignment; Kif4A and Kif4B (members of the kinesin-4 family) have roles in anaphase spindle dynamics; and Kif4A, Kif4B, MKLP1, and MKLP2 (members of the kinesin-6 family) are essential for cytokinesis. Using immunofluorescence analysis, time-lapse microscopy, and rescue experiments, we investigate the roles of these 12 kinesins in detail. INTRODUCTIONThe mitotic spindle, a unique cellular apparatus assembled at the beginning of mitosis, plays a pivotal role in regulating mitosis and cytokinesis. Although tubulins are the most abundant protein in the mitotic spindle, many additional proteins are involved in regulating its formation and function. Most prominent among these are members of the kinesin and dynein families of MT-based motor proteins (Mandelkow and Mandelkow, 2002;Vale, 2003), which generate directional movement along microtubules. The kinesin proteins share a conserved, ϳ340 amino acid, motor domain that utilizes ATP to fuel their movement along MTs. Outside the motor domain, kinesins also contain different "stalk" and "tail" domains that mediate oligomerization, regulation of motor activity, and interactions with their specific cargos (Vale, 2003).The roles of kinesins in mitosis and cytokinesis have been extensively studied in Saccharomyces cerevisiae, in which there are six kinesin genes belonging to five subfamilies. Gene disruption experiments have demonstrated that five of these kinesins play essential roles in regulating the formation, orientation, and elongation of the mitotic spindle and the segregation of chromosomes in mitosis (Hildebrandt and Hoyt, 2000). In addition, one cytoplasmic dynein motorcontaining polypeptide, the dynein heavy chain (DHC), has also been implicated in mitotic spindle function (Hildebrandt and Hoyt, 2000). Using RNAi techniques, Goshima and Vale recently examined the mitotic functions of cytoplasmic dynein and all 25 fly kinesins in cultured Drosophila cells (Goshima and Vale, 2003). They found that four kinesins are needed for bipolar spindle assembly and four are crucial for metaphase chromosome alignment. Dynein plays a role in the metaphase-to-anaphase transition, and one kinesin is required for cytokinesis.Th...

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Section: Discussioncontrasting

confidence: 53%

Functional Analysis of Human Microtubule-based Motor Proteins, the Kinesins and Dyneins, in Mitosis/Cytokinesis Using RNA Interference

Zhu

Zhao

Bibikova

et al. 2005

MBoC

374

344

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“…The action of Kif2A has been more controversial. Some studies implicate the protein in controlling MT flux (Gaetz and Kapoor, 2004;Ganem et al, 2005), whereas other studies clearly show that it is not required for MT flux (Cameron et al, 2006;Ohi et al, 2007). More recently, it has been demonstrated that although both MCAK and Kif2A can depolymerize MTs with nearly equal efficiency, they cannot substitute for each other functionally during spindle assembly in Xenopus egg extracts (Ohi et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

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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“…Studies in vertebrates and fruitflies have suggested that these proteins are transported poleward by cytoplasmic dynein. Inhibition of dynein in Xenopus egg extract spindles or Drosophila tissue culture cells causes kinesin-13s that are normally associated with poles to become spread across the spindle (Gaetz and Kapoor 2004;Morales-Mulia and Scholey 2005). We have also found that the accumulation of KLP10A on spindle poles involves a second kinesin-13 family member, KLP59D.…”

Section: Mechanism Of Flux I: Depolymerization Of Spindle Pole-focusementioning

confidence: 54%

The molecular basis of anaphase A in animal cells

Rath

Sharp

2011

Chromosome Res

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The mechanisms that move chromatids poleward during anaphase A have fascinated researchers for decades. There is now growing evidence that this movement is tightly linked to the active depolymerization of both ends of kinetochoreassociated microtubules, a mechanism we refer to as "Pacman-Flux." Contemporary data suggest that this is catalyzed by the integration of multiple enzymatic activities including (1) microtubule-end depolymerases housed at the pole or kinetochore, (2) microtubule-severing enzymes used to uncap the ends of kinetochore-associated microtubules, and (3) molecular motors which drive tubulins towards the pole or into kinetochores.

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Dynein/dynactin regulate metaphase spindle length by targeting depolymerizing activities to spindle poles

Cited by 184 publications

References 31 publications

Functional Analysis of Human Microtubule-based Motor Proteins, the Kinesins and Dyneins, in Mitosis/Cytokinesis Using RNA Interference

Functional Analysis of Human Microtubule-based Motor Proteins, the Kinesins and Dyneins, in Mitosis/Cytokinesis Using RNA Interference

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

The molecular basis of anaphase A in animal cells

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