Target-identification and mechanism-of-action studies of indole terpenoid mimics reveal that proper spindle microtubule assembly is essential for Cdh1-mediated proteolysis of CENP-A

Peng, Y.; Zhang, Y.; Fang, R.; Jiang, H.; Lan, G.; Liu, Y.; Nie, Z.; Zhang, S.-D.; Ma, Y.; Yang, P.; Wang, F.; Ge, H.-H.; Zhang, W.-D.; Luo, C.; Li, A.; He, W.

doi:10.1101/2023.04.25.538200

bioRxiv · 10.1101/2023.04.25.538200

Target-identification and mechanism-of-action studies of indole terpenoid mimics reveal that proper spindle microtubule assembly is essential for Cdh1-mediated proteolysis of CENP-A

Peng, Y.·Zhang, Y.·Fang, R.·Jiang, H.·Lan, G.·Liu, Y.·Nie, Z.·Zhang, S.-D.·Ma, Y.·Yang, P.·Wang, F.·Ge, H.-H.·Zhang, W.-D.·Luo, C.·Li, A.·He, W.

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Abstract

Centromere protein A (CENP-A), a histone H3 variant specific to centromeres, is crucial for kinetochore positioning and chromosome segregation. However, its regulatory mechanism in human cells remains incompletely understood. We conducted a structure-activity relationship (SAR) study of the cell cycle-arresting indole terpenoid mimic JP18 and found two more potent analogues, (+)-6-Br-JP18 and (+)-6-Cl-JP18. Tubulin was identified as a potential cellular target of these halogenated analogues by using the drug affinity responsive target stability (DARTS) based method. X-ray crystallography analysis revealed that both molecules bind to the colchicine-binding site of {beta}-tubulin. Furthermore, we discovered that treatment of human cells with microtubule-targeting agents (MTAs), including these two compounds, led to CENP-A accumulation by destabilizing Cdh1, a co-activator of the APC/C E3 ubiquitin ligase. Our study establishes a link between microtubule dynamics and CENP-A accumulation using small-molecule tools and highlights the role of Cdh1 in CENP-A proteolysis.

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Peng, Y., Zhang, Y., Fang, R., Jiang, H., Lan, G., Liu, Y., Nie, Z., Zhang, S.-D., Ma, Y., Yang, P., Wang, F., Ge, H.-H., Zhang, W.-D., Luo, C., Li, A., He, W.. 2023-04-26. Target-identification and mechanism-of-action studies of indole terpenoid mimics reveal that proper spindle microtubule assembly is essential for Cdh1-mediated proteolysis of CENP-A. https://doi.org/10.1101/2023.04.25.538200

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