Bergmann, Jan HRodríguez, Mariluz GómezMartins, Nuno M CKimura, HiroshiKelly, David AMasumoto, HiroshiLarionov, VladimirJansen, Lars E TEarnshaw, William C2016-05-242016-05-242011-01-19Bergmann, J. H., Rodríguez, M. G., Martins, N. M. C., Kimura, H., Kelly, D. A., Masumoto, H., Larionov, V., Jansen, L. E. T., Earnshaw, W. C. (2011). Epigenetic engineering shows H3K4me2 is required for HJURP targeting and CENP-A assembly on a synthetic human kinetochore. EMBO J., 30(2), 328–340.http://hdl.handle.net/10400.7/613Kinetochores assemble on distinct 'centrochromatin' containing the histone H3 variant CENP-A and interspersed nucleosomes dimethylated on H3K4 (H3K4me2). Little is known about how the chromatin environment at active centromeres governs centromeric structure and function. Here, we report that centrochromatin resembles K4-K36 domains found in the body of some actively transcribed housekeeping genes. By tethering the lysine-specific demethylase 1 (LSD1), we specifically depleted H3K4me2, a modification thought to have a role in transcriptional memory, from the kinetochore of a synthetic human artificial chromosome (HAC). H3K4me2 depletion caused kinetochores to suffer a rapid loss of transcription of the underlying α-satellite DNA and to no longer efficiently recruit HJURP, the CENP-A chaperone. Kinetochores depleted of H3K4me2 remained functional in the short term, but were defective in incorporation of CENP-A, and were gradually inactivated. Our data provide a functional link between the centromeric chromatin, α-satellite transcription, maintenance of CENP-A levels and kinetochore stability.engAutoantigensCentromereChromatinChromatin ImmunoprecipitationChromosomal Proteins, Non-HistoneChromosomes, Artificial, HumanDNA PrimersDNA-Binding ProteinsEpigenesis, GeneticGenetic EngineeringHistonesHumansKinetochoresNucleosomesReverse Transcriptase Polymerase Chain Reactionjournal article10.1038/emboj.2010.329