Stankovic, AnaGuo, Lucie Y.Mata, João F.Bodor, Dani L.Cao, Xing-JunBailey, Aaron O.Shabanowitz, JeffreyHunt, Donald F.Garcia, Benjamin A.Black, Ben E.Jansen, Lars E.T.2017-11-132018-01-192017-01-19Ana Stankovic, Lucie Y. Guo, João F. Mata, Dani L. Bodor, Xing-Jun Cao, Aaron O. Bailey, Jeffrey Shabanowitz, Donald F. Hunt, Benjamin A. Garcia, Ben E. Black, Lars E.T. Jansen, A Dual Inhibitory Mechanism Sufficient to Maintain Cell-Cycle-Restricted CENP-A Assembly, In Molecular Cell, Volume 65, Issue 2, 2017, Pages 231-246, ISSN 1097-2765, https://doi.org/10.1016/j.molcel.2016.11.021. (http://www.sciencedirect.com/science/article/pii/S1097276516307572)http://hdl.handle.net/10400.7/804The deposited article is a post-print version and has peer review. The deposited article version contains attached the supplementary materials within the pdf. This publication hasn't any creative commons license associated.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.engcentromerehistone variantCENP-Amitosisepigeneticschromatincyclin dependent kinasecell cyclekinetochoreA Dual Inhibitory Mechanism Sufficient to Maintain Cell-Cycle-Restricted CENP-A Assemblyjournal article10.1016/j.molcel.2016.11.021