TY - JOUR TI - Chromatin structure-dependent histone incorporation revealed by a genome-wide deposition assay AU - Tachiwana, Hiroaki AU - Dacher, Mariko AU - Maehara, Kazumitsu AU - Harada, Akihito AU - Seto, Yosuke AU - Katayama, Ryohei AU - Ohkawa, Yasuyuki AU - Kimura, Hiroshi AU - Kurumizaka, Hitoshi AU - Saitoh, Noriko A2 - Tyler, Jessica K A2 - Mattiroli, Francesca VL - 10 PY - 2021 DA - 2021/05/10 SP - e66290 C1 - eLife 2021;10:e66290 DO - 10.7554/eLife.66290 UR - https://doi.org/10.7554/eLife.66290 AB - In eukaryotes, histone variant distribution within the genome is the key epigenetic feature. To understand how each histone variant is targeted to the genome, we developed a new method, the RhIP (Reconstituted histone complex Incorporation into chromatin of Permeabilized cell) assay, in which epitope-tagged histone complexes are introduced into permeabilized cells and incorporated into their chromatin. Using this method, we found that H3.1 and H3.3 were incorporated into chromatin in replication-dependent and -independent manners, respectively. We further found that the incorporation of histones H2A and H2A.Z mainly occurred at less condensed chromatin (open), suggesting that condensed chromatin (closed) is a barrier for histone incorporation. To overcome this barrier, H2A, but not H2A.Z, uses a replication-coupled deposition mechanism. Our study revealed that the combination of chromatin structure and DNA replication dictates the differential histone deposition to maintain the epigenetic chromatin states. KW - chromatin KW - Histone KW - nucleosome KW - H2A.Z JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -