Molecular Interplay Between HURP and Kif18A in Mitotic Spindle Regulation

Overview

Journal Res Sq

Date 2024 Jun 10

PMID 38854046

Authors

Juan M Perez-Bertoldi

Yuanchang Zhao

Akanksha Thawani

Ahmet Yildiz

Eva Nogales

Affiliations

Soon will be listed here.

Abstract

During mitosis, microtubule dynamics are regulated to ensure proper alignment and segregation of chromosomes. The dynamics of kinetochore-attached microtubules are regulated by hepatoma-upregulated protein (HURP) and the mitotic kinesin-8 Kif18A, but the underlying mechanism remains elusive. Using single-molecule imaging , we demonstrate that Kif18A motility is regulated by HURP. While sparse decoration of HURP activates the motor, higher concentrations hinder processive motility. To shed light on this behavior, we determined the binding mode of HURP to microtubules using Cryo-EM. The structure reveals that one HURP motif spans laterally across β-tubulin, while a second motif binds between adjacent protofilaments. HURP partially overlaps with the microtubule-binding site of the Kif18A motor domain, indicating that excess HURP inhibits Kif18A motility by steric exclusion. We also observed that HURP and Kif18A function together to suppress dynamics of the microtubule plus-end, providing a mechanistic basis for how they collectively serve in spindle length control.

References

Zhang R, LaFrance B, Nogales E . Separating the effects of nucleotide and EB binding on microtubule structure. Proc Natl Acad Sci U S A. 2018; 115(27):E6191-E6200. PMC: 6142192. DOI: 10.1073/pnas.1802637115. View

Stumpff J, Du Y, English C, Maliga Z, Wagenbach M, Asbury C . A tethering mechanism controls the processivity and kinetochore-microtubule plus-end enrichment of the kinesin-8 Kif18A. Mol Cell. 2011; 43(5):764-75. PMC: 3172727. DOI: 10.1016/j.molcel.2011.07.022. View

Niwa S, Lipton D, Morikawa M, Zhao C, Hirokawa N, Lu H . Autoinhibition of a Neuronal Kinesin UNC-104/KIF1A Regulates the Size and Density of Synapses. Cell Rep. 2016; 16(8):2129-2141. PMC: 5432123. DOI: 10.1016/j.celrep.2016.07.043. View

Dudka D, Castrogiovanni C, Liaudet N, Vassal H, Meraldi P . Spindle-Length-Dependent HURP Localization Allows Centrosomes to Control Kinetochore-Fiber Plus-End Dynamics. Curr Biol. 2019; 29(21):3563-3578.e6. DOI: 10.1016/j.cub.2019.08.061. View

Espeut J, Gaussen A, Bieling P, Morin V, Prieto S, Fesquet D . Phosphorylation relieves autoinhibition of the kinetochore motor Cenp-E. Mol Cell. 2008; 29(5):637-43. DOI: 10.1016/j.molcel.2008.01.004. View

Jumper J, Evans R, Pritzel A, Green T, Figurnov M, Ronneberger O . Highly accurate protein structure prediction with AlphaFold. Nature. 2021; 596(7873):583-589. PMC: 8371605. DOI: 10.1038/s41586-021-03819-2. View

Hunter A, Caplow M, Coy D, Hancock W, Diez S, Wordeman L . The kinesin-related protein MCAK is a microtubule depolymerase that forms an ATP-hydrolyzing complex at microtubule ends. Mol Cell. 2003; 11(2):445-57. PMC: 6468321. DOI: 10.1016/s1097-2765(03)00049-2. View

Liebschner D, Afonine P, Baker M, Bunkoczi G, Chen V, Croll T . Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix. Acta Crystallogr D Struct Biol. 2019; 75(Pt 10):861-877. PMC: 6778852. DOI: 10.1107/S2059798319011471. View

Joglekar A, Hunt A . A simple, mechanistic model for directional instability during mitotic chromosome movements. Biophys J. 2002; 83(1):42-58. PMC: 1302126. DOI: 10.1016/S0006-3495(02)75148-5. View

10.

Tan Z, Yue Y, Leprevost F, Haynes S, Basrur V, Nesvizhskii A . Autoinhibited kinesin-1 adopts a hierarchical folding pattern. Elife. 2023; 12. PMC: 10619981. DOI: 10.7554/eLife.86776. View

11.

Potapova T, Gorbsky G . The Consequences of Chromosome Segregation Errors in Mitosis and Meiosis. Biology (Basel). 2017; 6(1). PMC: 5372005. DOI: 10.3390/biology6010012. View

12.

Hooikaas P, Martin M, Muhlethaler T, Kuijntjes G, Peeters C, Katrukha E . MAP7 family proteins regulate kinesin-1 recruitment and activation. J Cell Biol. 2019; 218(4):1298-1318. PMC: 6446838. DOI: 10.1083/jcb.201808065. View

13.

Song L, Craney A, Rape M . Microtubule-dependent regulation of mitotic protein degradation. Mol Cell. 2014; 53(2):179-92. PMC: 3926196. DOI: 10.1016/j.molcel.2013.12.022. View

14.

Zhang R, Roostalu J, Surrey T, Nogales E . Structural insight into TPX2-stimulated microtubule assembly. Elife. 2017; 6. PMC: 5679754. DOI: 10.7554/eLife.30959. View

15.

Stumpff J, Wagenbach M, Franck A, Asbury C, Wordeman L . Kif18A and chromokinesins confine centromere movements via microtubule growth suppression and spatial control of kinetochore tension. Dev Cell. 2012; 22(5):1017-29. PMC: 3356572. DOI: 10.1016/j.devcel.2012.02.013. View

16.

Santaguida S, Amon A . Short- and long-term effects of chromosome mis-segregation and aneuploidy. Nat Rev Mol Cell Biol. 2015; 16(8):473-85. DOI: 10.1038/nrm4025. View

17.

Hanks S, Coleman K, Reid S, Plaja A, Firth H, Fitzpatrick D . Constitutional aneuploidy and cancer predisposition caused by biallelic mutations in BUB1B. Nat Genet. 2004; 36(11):1159-61. DOI: 10.1038/ng1449. View

18.

Wu J, Chen C, Coumar M, Lin W, Chen Z, Hsu J . Aurora kinase inhibitors reveal mechanisms of HURP in nucleation of centrosomal and kinetochore microtubules. Proc Natl Acad Sci U S A. 2013; 110(19):E1779-87. PMC: 3651446. DOI: 10.1073/pnas.1220523110. View

19.

Du Y, English C, Ohi R . The kinesin-8 Kif18A dampens microtubule plus-end dynamics. Curr Biol. 2010; 20(4):374-80. DOI: 10.1016/j.cub.2009.12.049. View

20.

Mann B, Balchand S, Wadsworth P . Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics. Mol Biol Cell. 2016; 28(1):65-75. PMC: 5221630. DOI: 10.1091/mbc.E16-06-0476. View