The Microtubule-destabilizing Kinesin XKCM1 Regulates Microtubule Dynamic Instability in Cells

Kline-Smith, Susan L.; Walczak, Claire

doi:10.1091/mbc.e01-12-0143

Cited by 151 publications

(146 citation statements)

References 41 publications

Supporting

Mentioning

133

Contrasting

Unclassified

Order By: Relevance

“…The large increase in astral MTs seen after knockdown of MCAK is consistent with earlier studies showing that MCAK and its orthologues have a dramatic effect on astral MT organization (Walczak et al, 1996;Grill et al, 2001;Maney et al, 2001;Kline-Smith and Walczak, 2002;Goshima and Vale, 2003;Rogers et al, 2004;Srayko et al, 2005). Unfortunately, we were unable to obtain FRAP recovery data on the astral MTs because of the difficulty in obtaining sufficient fluorescence signal to get reliable FRAP curves.…”

Section: What Is the Relationship Between Spindle Mt Dynamics And Orgsupporting

confidence: 88%

“…Microinjection was carried out as described in Kline-Smith and Walczak (2002), by using either control nonimmune immunoglobulin G (IgG), antiXenopus MCAK (Walczak et al, 1996), or anti-PtK MCAK antibodies, each at needle concentration of 2 mg/ml. Antibodies were stored in antibody dialysis buffer (10 mM HEPES and 100 mM KCl, pH 7.2).…”

Section: Microinjectionmentioning

confidence: 99%

“…These include members of the kinesin-13 family (Kif2A, Kif2B, and MCAK/Kif2C), which seem to be strictly MT-depolymerizing enzymes (Desai et al, 1999;Hunter et al, 2003;Helenius et al, 2006), as well as members of the Kinesin-8 family, which are MT plus end-directed motors and plus end-specific depolymerases (Gupta et al, 2006;Howard and Hyman, 2007;Mayr et al, 2007;Varga et al, 2008). MCAK has been shown to be a critical regulator of MT dynamics and organization in vitro, in Xenopus egg extracts and in mammalian cells (Walczak et al, 1996;Desai et al, 1999;Maney et al, 2001;Kline-Smith and Walczak, 2002;Cassimeris and Morabito, 2004;Zhu et al, 2005;Stout et al, 2006;Manning et al, 2007;Ohi et al, 2007;Wordeman et al, 2007;Hedrick et al, 2008). In addition, the action of MCAK is critical at the kinetochore in which it may promote error correction by facilitating kinetochore MT turnover and the coordination of chromosome movement .…”

mentioning

confidence: 99%

See 2 more Smart Citations

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

Rizk

Bohannon

Wetzel

et al. 2009

MBoC

Self Cite

View full text Add to dashboard Cite

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...

show abstract

Section: What Is the Relationship Between Spindle Mt Dynamics And Orgsupporting

confidence: 88%

Section: Microinjectionmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

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

Rizk

Bohannon

Wetzel

et al. 2009

MBoC

Self Cite

View full text Add to dashboard Cite

show abstract

“…Mitotic centromere-associated kinesin overexpression causes a moderate increase in the frequency of multipolar spindles (Holmfeldt et al, 2004) and monopolar spindles (Kline-Smith and Walczak, 2002), which can contribute to the gain or loss of chromosomes in daughter cells. We reported that overexpression of MCAK leads to the increased migratory and proliferative ability of gastric cancer cells (Nakamura et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

“…Mitotic centromereassociated kinesin destabilises MTs from either end, and this activity and localisation are under the regulation of mitotic kinases (Wordeman et al, 2007). Overexpression of wild-type MCAK induces destabilisation of MTs in both mitotic and interphase cells (Kline-Smith and Walczak, 2002).…”

mentioning

confidence: 99%

Mitotic centromere-associated kinesin is a novel marker for prognosis and lymph node metastasis in colorectal cancer

et al. 2008

View full text Add to dashboard Cite

Mitotic centromere-associated kinesin (MCAK) is a microtubule depolymerase that is essential for proper kinetochore -microtubule attachment during spindle formation. Overexpression of MCAK has been correlated with aggressive forms of carcinoma, resulting in poor prognosis of colorectal cancer. The purpose of this study was to quantify MCAK expression in malignant and benign colorectal tissues and to determine if MCAK expression levels correlate with clinicopathologic factors and prognosis in colorectal cancer patients. Paired colorectal tissue samples from tumours and the corresponding normal tissues were obtained from 120 patients with colorectal cancer who underwent surgical resection. The real-time reverse transcriptase-PCR and immunohistochemistry were used to analyse mRNA and protein expression status with respect to various clinicopathological factors. MCAK expression was higher in colorectal cancer tissue (Po0.01) than in corresponding normal tissue, and this elevated expression level was markedly associated with factors such as lymph node metastasis (P ¼ 0.0023), venous invasion (P ¼ 0.019), peritoneal dissemination (P ¼ 0.021) and Dukes' classification (P ¼ 0.0023). Patients with high MCAK mRNA expression also showed a far poorer survival rate than those with low MCAK mRNA expression (Po0.01). Elevated MCAK expression was an independent predictor of overall survival and lymph node metastasis. These data suggest that MCAK expression may serve as a good marker of prognosis and lymph node metastasis in colorectal cancer.

show abstract

G protein βγ subunits interact with αβ‐ and γ‐tubulin and play a role in microtubule assembly in PC12 cells

Montoya

Gutierrez

Najera

et al. 2007

Cell Motil. Cytoskeleton

View full text Add to dashboard Cite

The betagamma subunit of G proteins (Gbetagamma) is known to transfer signals from cell surface receptors to intracellular effector molecules. Recent results suggest that Gbetagamma also interacts with microtubules and is involved in the regulation of the mitotic spindle. In the current study, the anti-microtubular drug nocodazole was employed to investigate the mechanism by which Gbetagamma interacts with tubulin and its possible implications in microtubule assembly in cultured PC12 cells. Nocodazole-induced depolymerization of microtubules drastically inhibited the interaction between Gbetagamma and tubulin. Gbetagamma was preferentially bound to microtubules and treatment with nocodazole suggested that the dissociation of Gbetagamma from microtubules is an early step in the depolymerization process. When microtubules were allowed to recover after removal of nocodazole, the tubulin-Gbetagamma interaction was restored. Unlike Gbetagamma, however, the interaction between tubulin and the alpha subunit of the Gs protein (Gsalpha) was not inhibited by nocodazole, indicating that the inhibition of tubulin-Gbetagamma interactions during microtubule depolymerization is selective. We found that Gbetagamma also interacts with gamma-tubulin, colocalizes with gamma-tubulin in centrosomes, and co-sediments in centrosomal fractions. The interaction between Gbetagamma and gamma-tubulin was unaffected by nocodazole, suggesting that the Gbetagamma-gamma-tubulin interaction is not dependent on assembled microtubules. Taken together, our results suggest that Gbetagamma may play an important and definitive role in microtubule assembly and/or stability. We propose that betagamma-microtubule interaction is an important step for G protein-mediated cell activation. These results may also provide new insights into the mechanism of action of anti-microtubule drugs.

show abstract

The Microtubule-destabilizing Kinesin XKCM1 Regulates Microtubule Dynamic Instability in Cells

Cited by 151 publications

References 41 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

Mitotic centromere-associated kinesin is a novel marker for prognosis and lymph node metastasis in colorectal cancer

G protein βγ subunits interact with αβ‐ and γ‐tubulin and play a role in microtubule assembly in PC12 cells

Contact Info

Product

Resources

About