Identification of a novel protein regulating microtubule stability through a chemical approach.
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.
Wignall
Sarah M
SM
Department of Molecular and Cell Biology, University of California, Berkeley, 311 Life Sciences Addition, Berkeley, California 94720, USA.
Gray
Nathanael S
NS
Chang
Young-Tae
YT
Juarez
Lolita
L
Jacob
Richard
R
Burlingame
Al
A
Schultz
Peter G
PG
Heald
Rebecca
R
D000595:Amino Acid Sequence; D000818:Animals; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002875:Chromosomes; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology; D015870:Gene Expression; D015723:Gene Library; D006801:Humans; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000502:physiology; D008870:Microtubules / Q000502:physiology*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010770:Phosphotransferases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011687:Purines / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology; D008941:Spindle Apparatus / Q000502:physiology; D013440:Sulfides / Q000302:isolation & purification* / Q000494:pharmacology; D014404:Tubulin / Q000378:metabolism; D050257:Tubulin Modulators; D014981:Xenopus
D004791:Enzyme Inhibitors; D008869:Microtubule-Associated Proteins; D011687:Purines; D013440:Sulfides; D014404:Tubulin; D050257:Tubulin Modulators; C491035:diminutol; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; D010770:Phosphotransferases