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.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Animals
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Cell Division / drug effects
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Cell Division / physiology
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Chromosomes
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Cytoskeleton / drug effects
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Cytoskeleton / physiology
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Enzyme Inhibitors / metabolism
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Enzyme Inhibitors / pharmacology
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Gene Expression
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Gene Library
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Humans
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Mice
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Microtubule-Associated Proteins / chemistry
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Microtubule-Associated Proteins / physiology
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Microtubules / physiology*
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Molecular Sequence Data
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Molecular Structure
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NAD(P)H Dehydrogenase (Quinone) / antagonists & inhibitors
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NAD(P)H Dehydrogenase (Quinone) / metabolism*
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Phosphotransferases / antagonists & inhibitors
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Phosphotransferases / metabolism
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Purines / analysis
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Purines / isolation & purification*
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Purines / pharmacology
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Spindle Apparatus / physiology
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Sulfides / isolation & purification*
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Sulfides / pharmacology
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Tubulin / metabolism
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Tubulin Modulators
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Xenopus
Substances
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Enzyme Inhibitors
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Microtubule-Associated Proteins
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Purines
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Sulfides
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Tubulin
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Tubulin Modulators
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diminutol
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NAD(P)H Dehydrogenase (Quinone)
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NQO1 protein, human
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Phosphotransferases