Mitotic Regulator SKAP Forms a Link between Kinetochore Core Complex KMN and Dynamic Spindle Microtubules

Wang, Xiwei; Zhuang, Xiaoxuan; Cao, Dan; Chu, Youjun; Yao, Phil; Liu, Wei; Liu, Lifang; Adams, Gregory; Fang, Guowei; Dou, Zhen; Ding, Xia; Huang, Yunpeng; Wang, Dongmei; Yao, Xuebiao

doi:10.1074/jbc.m112.406652

Cited by 47 publications

(81 citation statements)

References 43 publications

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“…Pulldown and Immunoprecipitation Assay-Pulldown assays were carried out as described previously (42). Briefly, the GSTtagged proteins were purified from bacteria by glutathione-agarose chromatography.…”

Section: Methodsmentioning

confidence: 99%

The Microtubule Plus End Tracking Protein TIP150 Interacts with Cortactin to Steer Directional Cell Migration

Adams

Zhang

Wang

et al. 2016

Journal of Biological Chemistry

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Cell migration is orchestrated by dynamic interactions of microtubules with the plasma membrane cortex. How these interactions facilitate these dynamic processes is still being actively investigated. TIP150 is a newly characterized microtubule plus end tracking protein essential for mitosis and entosis (Ward, T., Wang, M., Liu, X., Wang, Z., Xia, P., Chu, Y., Wang, X., Liu, L., Jiang, K., Yu, H., Yan, M., Wang, J., Hill, D. L., Huang, Y., Zhu, T., and Yao, X. (2013) Regulation of a dynamic interaction between two microtubule-binding proteins, EB1 and TIP150, by the mitotic p300/CBP-associated factor (PCAF) orchestrates kinetochore microtubule plasticity and chromosome stability during mitosis. J. Biol. Chem. 288, 15771-15785; Xia, P., Zhou, J., Song, X., Wu, B., Liu, X., Li, D., Zhang, S., Wang, Z., Yu, H., Ward, T., Zhang, J., Li, Y., Wang, X., Chen, Y., Guo, Z., and Yao, X. (2014) Aurora A orchestrates entosis by regulating a dynamic MCAK-TIP150 interaction. J. Mol. Cell Biol. 6, 240 -254). Here we show that TIP150 links dynamic microtubules to steer cell migration by interacting with cortactin. Mechanistically, TIP150 binds to cortactin via its C-terminal tail. Interestingly, the C-terminal TIP150 proline-rich region (CT150) binds to the Src homology 3 domain of cortactin specifically, and such an interaction is negatively regulated by EGF-elicited tyrosine phosphorylation of cortactin. Importantly, suppression of TIP150 or overexpression of phospho-mimicking cortactin inhibits polarized cell migration. In addition, CT150 disrupts the biochemical interaction between TIP150 and cortactin in vitro, and perturbation of the TIP150-cortactin interaction in vivo using a membrane-permeable TAT-CT150 peptide results in an inhibition of directional cell migration. We reason that a dynamic TIP150-cortactin interaction orchestrates directional cell migration via coupling dynamic microtubule plus ends to the cortical cytoskeleton.Cell migration is an essential physiological function that is tightly regulated in numerous biological processes, including development, tissue remodeling, wound healing, and tumor metastasis (1, 2). These processes require coordination of dynamic reorganization between the actin filaments and microtubules.Cortactin (CTN) 6 is a multidomain-containing scaffold protein that consists of an N-terminal acidic domain, six and a half tandem repeats, an ␣ helix, a proline-rich region, and an Src Homology (SH3) domain at its C terminus (3). CTN is an important scaffold for promoting the polymerization and rearrangement of the actin cytoskeleton (4). Functionally, CTN, distributed at protrusions and the leading edges, is necessary for cell migration by regulating intercellular adhesion and cell spreading (5). Importantly, CTN is phosphorylated at tyrosine residues in response to growth factor receptors, which regulates cell migration (6).Microtubules, one of the three major components of the cytoskeleton, are required for maintaining the physical and plastic properties of migrating cells (7). The plus en...

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Section: Methodsmentioning

confidence: 99%

The Microtubule Plus End Tracking Protein TIP150 Interacts with Cortactin to Steer Directional Cell Migration

Adams

Zhang

Wang

et al. 2016

Journal of Biological Chemistry

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“…In Vitro Pulldown Assay-Pulldown assays were carried out as described previously (21). Briefly, the GST fusion proteins in the soluble fraction were purified from bacteria by glutathioneagarose chromatography, whereas MBP-tagged proteins were purified using amylose beads and then eluted by MBP elution buffer (10 mM maltose in PBS).…”

Section: Methodsmentioning

confidence: 99%

“…Plasmids-GFP-and GST-tagged SKAP, full-length and truncations, were described previously (19,21). Bacterial expression constructs of SKAP full-length and truncations were ligated into the pMal-C2 vector (New England Biolabs, Beverly, MA).…”

Section: Methodsmentioning

confidence: 99%

“…To measure the EGF influence on SKAP distribution, MDA-MB-231 cells were starved over 6 -8 h, followed by treatment with EGF (100 ng/ml) for 15 min. Images were acquired with a DeltaVision wide-field deconvolution microscope (Applied Precision Inc., WA), as previously described (21).…”

Section: Methodsmentioning

confidence: 99%

“…Recently, we found that SKAP links kinetochore structural components to the spindle MTs through the Mis13-SKAP-CENP-E interaction pathway (19,21). Knocking down SKAP by siRNA is essential for accurate kinetochore-MT attachment.…”

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confidence: 99%

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Signaling Scaffold Protein IQGAP1 Interacts with Microtubule Plus-end Tracking Protein SKAP and Links Dynamic Microtubule Plus-end to Steer Cell Migration

Cao

Wang

et al. 2015

Journal of Biological Chemistry

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Background: IQGAP1 is a scaffold protein essential for cellular signaling in response to external cues. Results: Perturbation of the IQGAP1-SKAP interaction results in inhibition of directional cell migration. Conclusion: The IQGAP1-SKAP interaction links to dynamic microtubules at the leading edge to steer cell migration. Significance: IQGAP1 serves as a signaling hub for dynamic interaction between microtubule plus-ends and the cell cortex.

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Mapping the kinetochore MAP functions required for stabilizing microtubule attachments to chromosomes during metaphase

2019

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In mitosis, faithful chromosome segregation is orchestrated by the dynamic interactions between the spindle microtubules (MTs) emanating from the opposite poles and the kinetochores of the chromosomes. However, the precise mechanism that coordinates the coupling of the kinetochore components to dynamic MTs has been a long‐standing question. Microtubule‐associated proteins (MAPs) regulate MT nucleation and dynamics, MT‐mediated transport and MT cross‐linking in cells. During mitosis, MAPs play an essential role not only in determining spindle length, position, and orientation but also in facilitating robust kinetochore‐microtubule (kMT) attachments by linking the kinetochores to spindle MTs efficiently. The stability of MTs imparted by the MAPs is critical to ensure accurate chromosome segregation. This review primarily focuses on the specific function of nonmotor kinetochore MAPs, their recruitment to kinetochores and their MT‐binding properties. We also attempt to synthesize and strengthen our understanding of how these MAPs work in coordination with the kinetochore‐bound Ndc80 complex (the key component at the MT‐binding interface in metaphase and anaphase) to establish stable kMT attachments and control accurate chromosome segregation during mitosis.

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Mitotic Regulator SKAP Forms a Link between Kinetochore Core Complex KMN and Dynamic Spindle Microtubules

Cited by 47 publications

References 43 publications

The Microtubule Plus End Tracking Protein TIP150 Interacts with Cortactin to Steer Directional Cell Migration

The Microtubule Plus End Tracking Protein TIP150 Interacts with Cortactin to Steer Directional Cell Migration

Signaling Scaffold Protein IQGAP1 Interacts with Microtubule Plus-end Tracking Protein SKAP and Links Dynamic Microtubule Plus-end to Steer Cell Migration

Mapping the kinetochore MAP functions required for stabilizing microtubule attachments to chromosomes during metaphase

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