@article{670153e3382a48d686c17bb274481ae2,
title = "A Dual Inhibitory Mechanism Sufficient to Maintain Cell-Cycle-Restricted CENP-A Assembly",
abstract = "Chromatin featuring the H3 variant CENP-A at the centromere is critical for its mitotic function and epigenetic maintenance. Assembly of centromeric chromatin is restricted to G1 phase through inhibitory action of Cdk1/2 kinases in other phases of the cell cycle. Here, we identify the two key targets sufficient to maintain cell-cycle control of CENP-A assembly. We uncovered a single phosphorylation site in the licensing factor M18BP1 and a cyclin A binding site in the CENP-A chaperone, HJURP, that mediated specific inhibitory phosphorylation. Simultaneous expression of mutant proteins lacking these residues results in complete uncoupling from the cell cycle. Consequently, CENP-A assembly is fully recapitulated under high Cdk activities, indistinguishable from G1 assembly. We find that Cdk-mediated inhibition is exerted by sequestering active factors away from the centromere. Finally, we show that displacement of M18BP1 from the centromere is critical for the assembly mechanism of CENP-A.",
keywords = "CENP-A, cell cycle, centromere, chromatin, cyclin dependent kinase, epigenetics, histone variant, kinetochore, mitosis",
author = "Ana Stankovic and Guo, {Lucie Y.} and Mata, {Jo{\~a}o F.} and Bodor, {Dani L.} and Cao, {Xing Jun} and Bailey, {Aaron O.} and Jeffrey Shabanowitz and Hunt, {Donald F.} and Garcia, {Benjamin A.} and Black, {Ben E.} and Jansen, {Lars E.T.}",
note = "Funding Information: We thank N. Martins (Instituto Gulbenkian de Ci{\^e}ncia [IGC]) for preliminary work on HJURP mutants, E.V. Makeyev (Northwestern University [NU]), Singapore, and Dan Foltz (Northwestern University) for reagents, W. Siwek (IGC) for help with large-scale cell culture, S. Muller and G. Almouzni (Curie) for reagents and help with pre-extraction procedure, and T. Panchenko (Univeristy of Pennsylvania [UPenn]) for guidance with culturing LAP-tagged CENP-A cells and helpful discussions. We thank Monica Bettencourt Dias, Alekos Athanasiadis, Raquel Oliveira, and J{\"o}rg Becker (IGC) for critically reading the manuscript. Salary support to A.S. and D.L.B. was provided by Funda{\c c}{\~a}o para a Ci{\^e}ncia e a Tecnologia (FCT) fellowships SFRH/BD/51878/2012 and SFRH/BD/74284/2010, respectively, and an “Investigador FCT” position to L.E.T.J. This work is supported by NIH/National Institute of General Medical Sciences (NIGMS) grant R01-GM082989 (to B.E.B.), NIH/NIGMS grant T32-GM008275 (UPenn Structural Biology and Molecular Biophysics Training Grant T32-GM008275; to L.Y.G.), NIH/NCI grant F30-CA186430 (to L.Y.G.), NIHgrants GM 037537 (to D.F.H.) and GM 110174 (to B.A.G.). Work was further supported by the Funda{\c c}{\~a}o para a Ci{\^e}ncia e a Tecnologia (FCT) grant BIA-BCM/100557/2008 (to L.E.T.J.), EMBO installation grant 1818 (to L.E.T.J.), and ERC-consolidator grant ERC-2013-CoG-615638 (to L.E.T.J.). Publisher Copyright: {\textcopyright} 2017 Elsevier Inc.",
year = "2017",
month = jan,
day = "19",
doi = "10.1016/j.molcel.2016.11.021",
language = "English",
volume = "65",
pages = "231--246",
journal = "Molecular Cell",
issn = "1097-2765",
number = "2",
}