Disentangling the Molecular Determinants for Cenp-F Localization to Nuclear Pores and Kinetochores

Overview

Journal EMBO Rep

Specialty Molecular Biology

Date 2018 Apr 11

PMID 29632243

Citations 15

Authors

Alessandro Berto

Jinchao Yu

Stephanie Morchoisne-Bolhy

Chiara Bertipaglia

Richard Vallee

Julien Dumont

Francoise Ochsenbein

Raphael Guerois

Valerie Doye

Affiliations

Soon will be listed here.

Abstract

Cenp-F is a multifaceted protein implicated in cancer and developmental pathologies. The Cenp-F C-terminal region contains overlapping binding sites for numerous proteins that contribute to its functions throughout the cell cycle. Here, we focus on the nuclear pore protein Nup133 that interacts with Cenp-F both at nuclear pores in prophase and at kinetochores in mitosis, and on the kinase Bub1, known to contribute to Cenp-F targeting to kinetochores. By combining structural modeling and yeast two-hybrid assays, we generate an interaction model between a conserved helix within the Nup133 β-propeller and a short leucine zipper-containing dimeric segment of Cenp-F. We thereby create mutants affecting the Nup133/Cenp-F interface and show that they prevent Cenp-F localization to the nuclear envelope, but not to kinetochores. Conversely, a point mutation within an adjacent leucine zipper affecting the kinetochore targeting of Cenp-F KT-core domain impairs its interaction with Bub1, but not with Nup133, identifying Bub1 as the direct KT-core binding partner of Cenp-F. Finally, we show that Cenp-E redundantly contributes together with Bub1 to the recruitment of Cenp-F to kinetochores.

Citing Articles

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References

Moynihan K, Pooley R, Miller P, Kaverina I, Bader D . Murine CENP-F regulates centrosomal microtubule nucleation and interacts with Hook2 at the centrosome. Mol Biol Cell. 2009; 20(22):4790-803. PMC: 2777108. DOI: 10.1091/mbc.e09-07-0560. View

Musinipally V, Howes S, Alushin G, Nogales E . The microtubule binding properties of CENP-E's C-terminus and CENP-F. J Mol Biol. 2013; 425(22):4427-41. PMC: 3881371. DOI: 10.1016/j.jmb.2013.07.027. View

Laoukili J, Kooistra M, Bras A, Kauw J, Kerkhoven R, Morrison A . FoxM1 is required for execution of the mitotic programme and chromosome stability. Nat Cell Biol. 2005; 7(2):126-36. DOI: 10.1038/ncb1217. View

McKinley K, Cheeseman I . Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects. Dev Cell. 2017; 40(4):405-420.e2. PMC: 5345124. DOI: 10.1016/j.devcel.2017.01.012. View

Wood C, Bruning M, Ibarra A, Bartlett G, Thomson A, Sessions R . CCBuilder: an interactive web-based tool for building, designing and assessing coiled-coil protein assemblies. Bioinformatics. 2014; 30(21):3029-35. PMC: 4201159. DOI: 10.1093/bioinformatics/btu502. View

Ren Y, Seo H, Blobel G, Hoelz A . Structural and functional analysis of the interaction between the nucleoporin Nup98 and the mRNA export factor Rae1. Proc Natl Acad Sci U S A. 2010; 107(23):10406-11. PMC: 2890840. DOI: 10.1073/pnas.1005389107. View

Dong G, Fan H, Schneidman-Duhovny D, Webb B, Sali A . Optimized atomic statistical potentials: assessment of protein interfaces and loops. Bioinformatics. 2013; 29(24):3158-66. PMC: 3842762. DOI: 10.1093/bioinformatics/btt560. View

Soukoulis V, Reddy S, Pooley R, Feng Y, Walsh C, Bader D . Cytoplasmic LEK1 is a regulator of microtubule function through its interaction with the LIS1 pathway. Proc Natl Acad Sci U S A. 2005; 102(24):8549-54. PMC: 1150833. DOI: 10.1073/pnas.0502303102. View

Chan G, Schaar B, Yen T . Characterization of the kinetochore binding domain of CENP-E reveals interactions with the kinetochore proteins CENP-F and hBUBR1. J Cell Biol. 1998; 143(1):49-63. PMC: 2132809. DOI: 10.1083/jcb.143.1.49. View

10.

Ashar H, James L, Gray K, Carr D, Black S, Armstrong L . Farnesyl transferase inhibitors block the farnesylation of CENP-E and CENP-F and alter the association of CENP-E with the microtubules. J Biol Chem. 2000; 275(39):30451-7. DOI: 10.1074/jbc.M003469200. View

11.

Zhu X, Chang K, He D, Mancini M, Brinkley W, Lee W . The C terminus of mitosin is essential for its nuclear localization, centromere/kinetochore targeting, and dimerization. J Biol Chem. 1995; 270(33):19545-50. DOI: 10.1074/jbc.270.33.19545. View

12.

Evans H, Edwards L, Goodwin R . Conserved C-terminal domains of mCenp-F (LEK1) regulate subcellular localization and mitotic checkpoint delay. Exp Cell Res. 2007; 313(11):2427-37. PMC: 3991481. DOI: 10.1016/j.yexcr.2007.03.035. View

13.

Baffet A, Hu D, Vallee R . Cdk1 Activates Pre-mitotic Nuclear Envelope Dynein Recruitment and Apical Nuclear Migration in Neural Stem Cells. Dev Cell. 2015; 33(6):703-16. PMC: 4480218. DOI: 10.1016/j.devcel.2015.04.022. View

14.

Zilio N, Wehrkamp-Richter S, Boddy M . A new versatile system for rapid control of gene expression in the fission yeast Schizosaccharomyces pombe. Yeast. 2012; 29(10):425-34. PMC: 3478897. DOI: 10.1002/yea.2920. View

15.

Hu D, Baffet A, Nayak T, Akhmanova A, Doye V, Vallee R . Dynein recruitment to nuclear pores activates apical nuclear migration and mitotic entry in brain progenitor cells. Cell. 2013; 154(6):1300-13. PMC: 3822917. DOI: 10.1016/j.cell.2013.08.024. View

16.

Bilokapic S, Schwartz T . Structural and functional studies of the 252 kDa nucleoporin ELYS reveal distinct roles for its three tethered domains. Structure. 2013; 21(4):572-80. PMC: 4077343. DOI: 10.1016/j.str.2013.02.006. View

17.

Johnson P, McKnight S . Eukaryotic transcriptional regulatory proteins. Annu Rev Biochem. 1989; 58:799-839. DOI: 10.1146/annurev.bi.58.070189.004055. View

18.

Zuccolo M, Alves A, Galy V, Bolhy S, Formstecher E, Racine V . The human Nup107-160 nuclear pore subcomplex contributes to proper kinetochore functions. EMBO J. 2007; 26(7):1853-64. PMC: 1847668. DOI: 10.1038/sj.emboj.7601642. View

19.

Kim Y, Holland A, Lan W, Cleveland D . Aurora kinases and protein phosphatase 1 mediate chromosome congression through regulation of CENP-E. Cell. 2010; 142(3):444-55. PMC: 2921712. DOI: 10.1016/j.cell.2010.06.039. View

20.

Andreani J, Faure G, Guerois R . InterEvScore: a novel coarse-grained interface scoring function using a multi-body statistical potential coupled to evolution. Bioinformatics. 2013; 29(14):1742-9. DOI: 10.1093/bioinformatics/btt260. View