UNC-45A is a novel microtubule-associated protein and regulator of paclitaxel sensitivity in ovarian cancer cells

Ashley Mooneyham; Yoshie Iizuka; Qing Yang; Courtney Coombes; Mark McClellan; Vijayalakshmi Shridhar; Edith Emmings; Mihir Shetty; Liqiang Chen; Teng Ai; Joyce Meints; Michael K Lee; Melissa K Gardner; Martina Bazzaro

doi:10.1158/1541-7786.MCR-18-0670

UNC-45A is a novel microtubule-associated protein and regulator of paclitaxel sensitivity in ovarian cancer cells

Ashley Mooneyham, Yoshie Iizuka, Qing Yang, Courtney Coombes, Mark McClellan, Vijayalakshmi Shridhar, Edith Emmings, Mihir Shetty, Liqiang Chen, Teng Ai, Joyce Meints, Michael K Lee, Melissa K Gardner, Martina Bazzaro

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Abstract

UNC-45A, a highly conserved member of the UCS (UNC45A/CRO1/SHE4P) protein family of cochaperones, plays an important role in regulating cytoskeletal-associated functions in invertebrates and mammalian cells, including cytokinesis, exocytosis, cell motility, and neuronal development. Here, for the first time, UNC-45A is demonstrated to function as a mitotic spindle-associated protein that destabilizes microtubules (MT) activity. Using in vitro biophysical reconstitution and total internal reflection fluorescence microscopy analysis, we reveal that UNC-45A directly binds to taxol-stabilized MTs in the absence of any additional cellular cofactors or other MT-associated proteins and acts as an ATP-independent MT destabilizer. In cells, UNC-45A binds to and destabilizes mitotic spindles, and its depletion causes severe defects in chromosome congression and segregation. UNC-45A is overexpressed in human clinical specimens from chemoresistant ovarian cancer and that UNC-45A-overexpressing cells resist chromosome missegregation and aneuploidy when treated with clinically relevant concentrations of paclitaxel. Lastly, UNC-45A depletion exacerbates paclitaxel-mediated stabilizing effects on mitotic spindles and restores sensitivity to paclitaxel. Implications: These findings reveal novel and significant roles for UNC-45A in regulation of cytoskeletal dynamics, broadening our understanding of the basic mechanisms regulating MT stability and human cancer susceptibility to paclitaxel, one of the most widely used chemotherapy agents for the treatment of human cancers.

Original language	English (US)
Pages (from-to)	370-383
Number of pages	14
Journal	Molecular Cancer Research
Volume	17
Issue number	2
DOIs	https://doi.org/10.1158/1541-7786.MCR-18-0670
State	Published - Feb 2019

Bibliographical note

Funding Information:
The authors are grateful toDr. Henry Epstein (1944-2013) for his friendship and helpful discussions and to Dr. David Odde for providing helpful insights. We thank Guillermo Marques (University of Minnesota Imaging Center) for assistance with image analysis. This work was supported by Department of Defense Ovarian Cancer Research Program Grant OC160377, the Minnesota Ovarian Cancer Alliance, and the Randy Shaver Cancer Research Funds to M. Bazzaro. M. Gardner was supported by NIH grant NIGMS R01 GM-103833. A. Mooneyham was funded by the NIH T32 CA009138 training grant. The funders had no role in study design, data collection and analysis, and decision to publish or preparation of the manuscript.

Funding Information:
The authors are grateful to Dr. Henry Epstein (1944–2013) for his friendship and helpful discussions and to Dr. David Odde for providing helpful insights. We thank Guillermo Marques (University of Minnesota Imaging Center) for assistance with image analysis. This work was supported by Department of Defense Ovarian Cancer Research Program Grant OC160377, the Minnesota Ovarian Cancer Alliance, and the Randy Shaver Cancer Research Funds to M. Bazzaro. M. Gardner was supported by NIH grant NIGMS R01 GM-103833. A. Mooneyham was funded by the NIH T32 CA009138 training grant. The funders had no role in study design, data collection and analysis, and decision to publish or preparation of the manuscript.

Publisher Copyright:
© 2018 American Association for Cancer Research.

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Mooneyham, A., Iizuka, Y., Yang, Q., Coombes, C., McClellan, M., Shridhar, V., Emmings, E., Shetty, M., Chen, L., Ai, T., Meints, J., Lee, M. K., Gardner, M. K., & Bazzaro, M. (2019). UNC-45A is a novel microtubule-associated protein and regulator of paclitaxel sensitivity in ovarian cancer cells. Molecular Cancer Research, 17(2), 370-383. https://doi.org/10.1158/1541-7786.MCR-18-0670

Mooneyham, A, Iizuka, Y, Yang, Q, Coombes, C, McClellan, M, Shridhar, V, Emmings, E, Shetty, M, Chen, L, Ai, T, Meints, J, Lee, MK , Gardner, MK & Bazzaro, M 2019, 'UNC-45A is a novel microtubule-associated protein and regulator of paclitaxel sensitivity in ovarian cancer cells', Molecular Cancer Research, vol. 17, no. 2, pp. 370-383. https://doi.org/10.1158/1541-7786.MCR-18-0670

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title = "UNC-45A is a novel microtubule-associated protein and regulator of paclitaxel sensitivity in ovarian cancer cells",

abstract = "UNC-45A, a highly conserved member of the UCS (UNC45A/CRO1/SHE4P) protein family of cochaperones, plays an important role in regulating cytoskeletal-associated functions in invertebrates and mammalian cells, including cytokinesis, exocytosis, cell motility, and neuronal development. Here, for the first time, UNC-45A is demonstrated to function as a mitotic spindle-associated protein that destabilizes microtubules (MT) activity. Using in vitro biophysical reconstitution and total internal reflection fluorescence microscopy analysis, we reveal that UNC-45A directly binds to taxol-stabilized MTs in the absence of any additional cellular cofactors or other MT-associated proteins and acts as an ATP-independent MT destabilizer. In cells, UNC-45A binds to and destabilizes mitotic spindles, and its depletion causes severe defects in chromosome congression and segregation. UNC-45A is overexpressed in human clinical specimens from chemoresistant ovarian cancer and that UNC-45A-overexpressing cells resist chromosome missegregation and aneuploidy when treated with clinically relevant concentrations of paclitaxel. Lastly, UNC-45A depletion exacerbates paclitaxel-mediated stabilizing effects on mitotic spindles and restores sensitivity to paclitaxel. Implications: These findings reveal novel and significant roles for UNC-45A in regulation of cytoskeletal dynamics, broadening our understanding of the basic mechanisms regulating MT stability and human cancer susceptibility to paclitaxel, one of the most widely used chemotherapy agents for the treatment of human cancers.",

author = "Ashley Mooneyham and Yoshie Iizuka and Qing Yang and Courtney Coombes and Mark McClellan and Vijayalakshmi Shridhar and Edith Emmings and Mihir Shetty and Liqiang Chen and Teng Ai and Joyce Meints and Lee, {Michael K} and Gardner, {Melissa K} and Martina Bazzaro",

note = "Funding Information: The authors are grateful toDr. Henry Epstein (1944-2013) for his friendship and helpful discussions and to Dr. David Odde for providing helpful insights. We thank Guillermo Marques (University of Minnesota Imaging Center) for assistance with image analysis. This work was supported by Department of Defense Ovarian Cancer Research Program Grant OC160377, the Minnesota Ovarian Cancer Alliance, and the Randy Shaver Cancer Research Funds to M. Bazzaro. M. Gardner was supported by NIH grant NIGMS R01 GM-103833. A. Mooneyham was funded by the NIH T32 CA009138 training grant. The funders had no role in study design, data collection and analysis, and decision to publish or preparation of the manuscript. Funding Information: The authors are grateful to Dr. Henry Epstein (1944–2013) for his friendship and helpful discussions and to Dr. David Odde for providing helpful insights. We thank Guillermo Marques (University of Minnesota Imaging Center) for assistance with image analysis. This work was supported by Department of Defense Ovarian Cancer Research Program Grant OC160377, the Minnesota Ovarian Cancer Alliance, and the Randy Shaver Cancer Research Funds to M. Bazzaro. M. Gardner was supported by NIH grant NIGMS R01 GM-103833. A. Mooneyham was funded by the NIH T32 CA009138 training grant. The funders had no role in study design, data collection and analysis, and decision to publish or preparation of the manuscript. Publisher Copyright: {\textcopyright} 2018 American Association for Cancer Research.",

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AU - Yang, Qing

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AU - McClellan, Mark

AU - Shridhar, Vijayalakshmi

AU - Emmings, Edith

AU - Shetty, Mihir

AU - Chen, Liqiang

AU - Ai, Teng

AU - Meints, Joyce

AU - Lee, Michael K

AU - Gardner, Melissa K

AU - Bazzaro, Martina

N1 - Funding Information: The authors are grateful toDr. Henry Epstein (1944-2013) for his friendship and helpful discussions and to Dr. David Odde for providing helpful insights. We thank Guillermo Marques (University of Minnesota Imaging Center) for assistance with image analysis. This work was supported by Department of Defense Ovarian Cancer Research Program Grant OC160377, the Minnesota Ovarian Cancer Alliance, and the Randy Shaver Cancer Research Funds to M. Bazzaro. M. Gardner was supported by NIH grant NIGMS R01 GM-103833. A. Mooneyham was funded by the NIH T32 CA009138 training grant. The funders had no role in study design, data collection and analysis, and decision to publish or preparation of the manuscript. Funding Information: The authors are grateful to Dr. Henry Epstein (1944–2013) for his friendship and helpful discussions and to Dr. David Odde for providing helpful insights. We thank Guillermo Marques (University of Minnesota Imaging Center) for assistance with image analysis. This work was supported by Department of Defense Ovarian Cancer Research Program Grant OC160377, the Minnesota Ovarian Cancer Alliance, and the Randy Shaver Cancer Research Funds to M. Bazzaro. M. Gardner was supported by NIH grant NIGMS R01 GM-103833. A. Mooneyham was funded by the NIH T32 CA009138 training grant. The funders had no role in study design, data collection and analysis, and decision to publish or preparation of the manuscript. Publisher Copyright: © 2018 American Association for Cancer Research.

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N2 - UNC-45A, a highly conserved member of the UCS (UNC45A/CRO1/SHE4P) protein family of cochaperones, plays an important role in regulating cytoskeletal-associated functions in invertebrates and mammalian cells, including cytokinesis, exocytosis, cell motility, and neuronal development. Here, for the first time, UNC-45A is demonstrated to function as a mitotic spindle-associated protein that destabilizes microtubules (MT) activity. Using in vitro biophysical reconstitution and total internal reflection fluorescence microscopy analysis, we reveal that UNC-45A directly binds to taxol-stabilized MTs in the absence of any additional cellular cofactors or other MT-associated proteins and acts as an ATP-independent MT destabilizer. In cells, UNC-45A binds to and destabilizes mitotic spindles, and its depletion causes severe defects in chromosome congression and segregation. UNC-45A is overexpressed in human clinical specimens from chemoresistant ovarian cancer and that UNC-45A-overexpressing cells resist chromosome missegregation and aneuploidy when treated with clinically relevant concentrations of paclitaxel. Lastly, UNC-45A depletion exacerbates paclitaxel-mediated stabilizing effects on mitotic spindles and restores sensitivity to paclitaxel. Implications: These findings reveal novel and significant roles for UNC-45A in regulation of cytoskeletal dynamics, broadening our understanding of the basic mechanisms regulating MT stability and human cancer susceptibility to paclitaxel, one of the most widely used chemotherapy agents for the treatment of human cancers.

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UNC-45A is a novel microtubule-associated protein and regulator of paclitaxel sensitivity in ovarian cancer cells

Abstract

Bibliographical note

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UNC-45A is a novel microtubule-associated protein and regulator of paclitaxel sensitivity in ovarian cancer cells

Abstract

Bibliographical note

UN SDGs

Access

OpenUrl availability

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Fingerprint

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Olympus Fluoview FV1000 BX2 Upright Confocal

University Imaging Centers

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