Please use this identifier to cite or link to this item:
Title: Multisite phosphorylation of C-Nap1 releases it from Cep135 to trigger centrosome disjunction
Authors: Hardy, Tara
Lee, Miseon
Hames, Rebecca S.
Prosser, Suzanna L.
Cheary, Donna-Marie
Samant, Mugdha D.
Schultz, Francisca
Baxter, Joanne E.
Rhee, Kunsoo
Fry, Andrew M.
First Published: 2-Apr-2014
Publisher: The Company of Biologists Ltd
Citation: Journal of Cell Science, 2014, 127, pp. 2493-2506
Abstract: During mitotic entry, centrosomes separate to establish the bipolar spindle. Delays in centrosome separation can perturb chromosome segregation and promote genetic instability. However, interphase centrosomes are physically tethered by a proteinaceous linker composed of C-Nap1 (also known as CEP250) and the filamentous protein rootletin. Linker disassembly occurs at the onset of mitosis in a process known as centrosome disjunction and is triggered by the Nek2-dependent phosphorylation of C-Nap1. However, the mechanistic consequences of C-Nap1 phosphorylation are unknown. Here, we demonstrate that Nek2 phosphorylates multiple residues within the C-terminal domain of C-Nap1 and, collectively, these phosphorylation events lead to loss of oligomerization and centrosome association. Mutations in non-phosphorylatable residues that make the domain more acidic are sufficient to release C-Nap1 from the centrosome, suggesting that it is an increase in overall negative charge that is required for this process. Importantly, phosphorylation of C-Nap1 also perturbs interaction with the core centriolar protein, Cep135, and interaction of endogenous C-Nap1 and Cep135 proteins is specifically lost in mitosis. We therefore propose that multisite phosphorylation of C-Nap1 by Nek2 perturbs both oligomerization and Cep135 interaction, and this precipitates centrosome disjunction at the onset of mitosis.
DOI Link: 10.1242/jcs.142331
ISSN: 0021-9533
eISSN: 1477-9137
Version: Publisher Version
Status: Peer-reviewed
Type: Journal Article
Rights: Copyright © the authors, 2014. This is an open-access article distributed under the terms of the Creative Commons Attribution License ( ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Description: PMCID: PMC4038944
Appears in Collections:Published Articles, Dept. of Biochemistry

Files in This Item:
File Description SizeFormat 
J Cell Sci-2014-Hardy-2493-506.pdfPublished (publisher PDF)4.35 MBUnknownView/Open

Items in LRA are protected by copyright, with all rights reserved, unless otherwise indicated.