The small molecule CS1 inhibits mitosis and sister chromatid resolution in HeLa cells

• Published in: Biochimica et biophysica acta Vol. 1862; no. 5; pp. 1134 - 1147
• Main Authors: Wu, Xingkang, Li, Zhenyu, Shen, Yuemao
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.05.2018
• Subjects: CS1; Histone phosphorylation; Microtubule inhibitor; Mitotic arrest; Sister chromatid resolution; Spindle assembly checkpoint; CS1; Histone phosphorylation; Mitotic arrest; Sister chromatid resolution; Spindle assembly checkpoint; Microtubule inhibitor
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2018.01.012

Mitosis, the most dramatic event in the cell cycle, involves the reorganization of virtually all cellular components. Antimitotic agents are useful for dissecting the mechanism of this reorganization. Previously, we found that the small molecule CS1 accumulates cells in G2/M phase [1], but the mechanism of its action remains unknown. Cell cycle analysis, live cell imaging and nuclear staining were used. Chromosomal morphology was detected by chromosome spreading. The effects of CS1 on microtubules were confirmed by tubulin polymerization, colchicine tubulin-binding, cellular tubulin polymerization and immunofluorescence assays and by analysis of microtubule dynamics and molecular modeling. Histone phosphoproteomics was performed using mass spectrometry. Cell signaling cascades were analyzed using immunofluorescence, immunoprecipitation, immunoblotting, siRNA knockdown and chemical inhibition of specific proteins. The small molecule CS1 was shown to be an antimitotic agent. CS1 potently inhibited microtubule polymerization via interaction with the colchicine-binding pocket of tubulin in vitro and inhibited the formation of the spindle apparatus by reducing the bulk of growing microtubules in HeLa cells, which led to activation of the spindle assembly checkpoint (SAC) and mitotic arrest of HeLa cells. Compared with colchicine, CS1 impaired the progression of sister chromatid resolution independent of cohesin dissociation, and this was reversed by the removal of CS1. Additionally, CS1 induced unique histone phosphorylation patterns distinct from those induced by colchicine. CS1 is a unique antimitotic small molecule and a powerful tool with unprecedented value over colchicine that makes it possible to specifically and conditionally perturb mitotic progression. [Display omitted] •CS1 inhibits microtubule polymerization by binding to the colchicine-binding pocket of tubulin.•CS1 reversibly inhibits mitosis and sister chromatid resolution.•CS1 induces unique histone phosphorylation patterns that differ from those induced by colchicine.