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PMID: 18455984 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Implications for kinetochore-microtubule attachment from the structure of an engineered Ndc80 complex.

Cell ·Vol. 133 ·No. 3 ·2008-05-02 ·Pages 427-39

Ciferri C, Pasqualato S, Screpanti E, Varetti G, Santaguida S, Dos Reis G, Maiolica A, Polka J, De Luca JG, De Wulf P, Salek M, Rappsilber J, Moores CA, Salmon ED, Musacchio A

Abstract

Kinetochores are proteinaceous assemblies that mediate the interaction of chromosomes with the mitotic spindle. The 180 kDa Ndc80 complex is a direct point of contact between kinetochores and microtubules. Its four subunits contain coiled coils and form an elongated rod structure with functional globular domains at either end. We crystallized an engineered "bonsai" Ndc80 complex containing a shortened rod domain but retaining the globular domains required for kinetochore localization and microtubule binding. The structure reveals a microtubule-binding interface containing a pair of tightly interacting calponin-homology (CH) domains with a previously unknown arrangement. The interaction with microtubules is cooperative and predominantly electrostatic. It involves positive charges in the CH domains and in the N-terminal tail of the Ndc80 subunit and negative charges in tubulin C-terminal tails and is regulated by the Aurora B kinase. We discuss our results with reference to current models of kinetochore-microtubule attachment and centromere organization.

MeSH Terms
Amino Acid Sequence Cell Cycle Proteins/chemistry,genetics,metabolism Crystallography, X-Ray Cytoskeletal Proteins Humans Kinetochores/metabolism Mass Spectrometry Microtubule-Associated Proteins/chemistry,genetics,metabolism Microtubules/metabolism Models, Molecular Molecular Sequence Data Nuclear Proteins/chemistry,genetics,metabolism Protein Engineering Spindle Apparatus/metabolism
Chemicals
Cell Cycle Proteins Cytoskeletal Proteins Microtubule-Associated Proteins NDC80 protein, human NUF2 protein, human Nuclear Proteins SPC24 protein, human
Authors & Affiliations
15 authors, click to expand affiliations / ORCID
Ciferri Claudio
Department of Experimental Oncology, European Institute of Oncology, Via Adamello 16, I 20139 Milan, Italy.
Pasqualato Sebastiano
Screpanti Emanuela
Varetti Gianluca
Santaguida Stefano
Dos Reis Gabriel
Maiolica Alessio
Polka Jessica
De Luca Jennifer G
De Wulf Peter
Salek Mogjiborahman
Rappsilber Juri
Moores Carolyn A
Salmon Edward D
Musacchio Andrea
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Article Info
Journal
Cell
Abbr.
Cell
ISSN
1097-4172
DOI
10.1016/j.cell.2008.03.020
Published
2008-05-02
Pages
427-39
Language
English
Region
United States
NLM ID
0413066
PMCID
PMC4754795
Subset
IM
Grants
Telethon · GGP06044 · Italy
NCI NIH HHS · K01 CA125051 · United States
NIGMS NIH HHS · R37 GM024364 · United States
Databases
PDB
2VE7
Corrections
CommentIn
PMID: 18547515
External Links
PubMed source DOI: 10.1016/j.cell.2008.03.020 PMC Full Text View journal details
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