Identification of a Novel Protein Regulating Microtubule Stability through a Chemical Approach

Sarah M. Wignall; Nathanael S. Gray; Young Tae Chang; Lolita Juarez; Richard Jacob; A. Burlingame; Peter G. Schultz; Rebecca Heald

doi:10.1016/j.chembiol.2003.12.019

Identification of a Novel Protein Regulating Microtubule Stability through a Chemical Approach

Sarah M. Wignall, Nathanael S. Gray, Young Tae Chang, Lolita Juarez, Richard Jacob, A. Burlingame, Peter G. Schultz, Rebecca Heald^*

^*Corresponding author for this work

Molecular Biosciences

Research output: Contribution to journal › Article › peer-review

61 Scopus citations

Abstract

To identify novel proteins regulating the microtubule cytoskeleton, we screened a library of purine derivatives using mitotic spindle assembly in Xenopus egg extracts as an assay. Out of a collection of 1561 compounds, we identified 15 that destabilized microtubules without targeting tubulin directly, resulting in small spindles. Affinity chromatography with one compound, named diminutol, revealed a potential target as NQO1, an NADP-dependent oxidoreductase. A role for NQO1 in influencing microtubule polymerization was confirmed through inhibition studies using known inhibitors and immunodepletion. Therefore, this chemical approach has identified a novel factor required for microtubule morphogenesis and cell division.

Original language	English (US)
Pages (from-to)	135-146
Number of pages	12
Journal	Chemistry and Biology
Volume	11
Issue number	1
DOIs	https://doi.org/10.1016/j.chembiol.2003.12.019
State	Published - Jan 2004

Funding

We thank Shiuan Chen and Kebin Wu (Beckman Institute of the City of Hope, Duarte, CA) for measuring the K i of diminutol using human NQO1, Jennifer Banks for critical reading of the manuscript, the Heald lab for helpful discussions, and G.O. Nads for sperm nuclei. This work was supported by the National Institutes of Health (R.H., GM57839), the Pew Charitable Trust (R.H.), the Cancer Research Coordinating Committee (R.H.), and a predoctoral NSF fellowship (S.M.W.).

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Pharmacology
Drug Discovery
Clinical Biochemistry

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title = "Identification of a Novel Protein Regulating Microtubule Stability through a Chemical Approach",

abstract = "To identify novel proteins regulating the microtubule cytoskeleton, we screened a library of purine derivatives using mitotic spindle assembly in Xenopus egg extracts as an assay. Out of a collection of 1561 compounds, we identified 15 that destabilized microtubules without targeting tubulin directly, resulting in small spindles. Affinity chromatography with one compound, named diminutol, revealed a potential target as NQO1, an NADP-dependent oxidoreductase. A role for NQO1 in influencing microtubule polymerization was confirmed through inhibition studies using known inhibitors and immunodepletion. Therefore, this chemical approach has identified a novel factor required for microtubule morphogenesis and cell division.",

author = "Wignall, {Sarah M.} and Gray, {Nathanael S.} and Chang, {Young Tae} and Lolita Juarez and Richard Jacob and A. Burlingame and Schultz, {Peter G.} and Rebecca Heald",

note = "Funding Information: We thank Shiuan Chen and Kebin Wu (Beckman Institute of the City of Hope, Duarte, CA) for measuring the K i of diminutol using human NQO1, Jennifer Banks for critical reading of the manuscript, the Heald lab for helpful discussions, and G.O. Nads for sperm nuclei. This work was supported by the National Institutes of Health (R.H., GM57839), the Pew Charitable Trust (R.H.), the Cancer Research Coordinating Committee (R.H.), and a predoctoral NSF fellowship (S.M.W.).",

year = "2004",

month = jan,

doi = "10.1016/j.chembiol.2003.12.019",

language = "English (US)",

volume = "11",

pages = "135--146",

journal = "Chemistry and Biology",

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AU - Wignall, Sarah M.

AU - Gray, Nathanael S.

AU - Chang, Young Tae

AU - Juarez, Lolita

AU - Jacob, Richard

AU - Burlingame, A.

AU - Schultz, Peter G.

AU - Heald, Rebecca

N1 - Funding Information: We thank Shiuan Chen and Kebin Wu (Beckman Institute of the City of Hope, Duarte, CA) for measuring the K i of diminutol using human NQO1, Jennifer Banks for critical reading of the manuscript, the Heald lab for helpful discussions, and G.O. Nads for sperm nuclei. This work was supported by the National Institutes of Health (R.H., GM57839), the Pew Charitable Trust (R.H.), the Cancer Research Coordinating Committee (R.H.), and a predoctoral NSF fellowship (S.M.W.).

PY - 2004/1

Y1 - 2004/1

N2 - To identify novel proteins regulating the microtubule cytoskeleton, we screened a library of purine derivatives using mitotic spindle assembly in Xenopus egg extracts as an assay. Out of a collection of 1561 compounds, we identified 15 that destabilized microtubules without targeting tubulin directly, resulting in small spindles. Affinity chromatography with one compound, named diminutol, revealed a potential target as NQO1, an NADP-dependent oxidoreductase. A role for NQO1 in influencing microtubule polymerization was confirmed through inhibition studies using known inhibitors and immunodepletion. Therefore, this chemical approach has identified a novel factor required for microtubule morphogenesis and cell division.

AB - To identify novel proteins regulating the microtubule cytoskeleton, we screened a library of purine derivatives using mitotic spindle assembly in Xenopus egg extracts as an assay. Out of a collection of 1561 compounds, we identified 15 that destabilized microtubules without targeting tubulin directly, resulting in small spindles. Affinity chromatography with one compound, named diminutol, revealed a potential target as NQO1, an NADP-dependent oxidoreductase. A role for NQO1 in influencing microtubule polymerization was confirmed through inhibition studies using known inhibitors and immunodepletion. Therefore, this chemical approach has identified a novel factor required for microtubule morphogenesis and cell division.

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Identification of a Novel Protein Regulating Microtubule Stability through a Chemical Approach

Abstract

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ASJC Scopus subject areas

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