Targeting the trypanosome kinetochore with CLK1 protein kinase inhibitors

Saldivia, M. et al. (2020) Targeting the trypanosome kinetochore with CLK1 protein kinase inhibitors. Nature Microbiology, 5(10), pp. 1207-1216. (doi: 10.1038/s41564-020-0745-6) (PMID:32661312) (PMCID:PMC7610364)

Abstract

The kinetochore is a macromolecular structure that assembles on the centromeres of chromosomes and provides the major attachment point for spindle microtubules during mitosis. In Trypanosoma brucei, the proteins that make up the kinetochore are highly divergent; the inner kinetochore comprises at least 20 distinct and essential proteins (KKT1-20) that include four protein kinases-CLK1 (also known as KKT10), CLK2 (also known as KKT19), KKT2 and KKT3. Here, we report the identification and characterization of the amidobenzimidazoles (AB) protein kinase inhibitors that show nanomolar potency against T. brucei bloodstream forms, Leishmania and Trypanosoma cruzi. We performed target deconvolution analysis using a selection of 29 T. brucei mutants that overexpress known essential protein kinases, and identified CLK1 as a primary target. Biochemical studies and the co-crystal structure of CLK1 in complex with AB1 show that the irreversible competitive inhibition of CLK1 is dependent on a Michael acceptor forming an irreversible bond with Cys 215 in the ATP-binding pocket, a residue that is not present in human CLK1, thereby providing selectivity. Chemical inhibition of CLK1 impairs inner kinetochore recruitment and compromises cell-cycle progression, leading to cell death. This research highlights a unique drug target for trypanosomatid parasitic protozoa and a new chemical tool for investigating the function of their divergent kinetochores.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Paape, Dr Daniel and Ritchie, Mr Ryan and McCulloch, Professor Richard and Barrett, Professor Michael and Mottram, Professor Jeremy
• Authors: Saldivia, M., Fang, E., Ma, X., Myburgh, E., Carnielli, J. B.T., Bower-Lepts, C., Brown, E., Ritchie, R., Lakshminarayana, S. B., Chen, Y.-L., Patra, D., Ornelas, E., Koh, H. X.Y., Williams, S. L., Supek, F., Paape, D., McCulloch, R., Kaiser, M., Barrett, M. P., Jiricek, J., Diagana, T. T., Mottram, J. C., and Rao, S. P.S.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: Nature Microbiology
• Publisher: Nature Research
• ISSN: 2058-5276
• ISSN (Online): 2058-5276
• Published Online: 13 January 2020
• Copyright Holders: Copyright © 2020 The Authors
• First Published: First published in Nature Microbiology 5(10): 1207-1216
• Publisher Policy: Reproduced in accordance with the publisher copyright policy

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
170547		The Wellcome Centre for Molecular Parasitology ( Core Support )	Andrew Waters	Wellcome Trust (WELLCOTR)	104111/Z/14/Z	III - Parasitology