RT Journal Article
SR Electronic
T1 Centromeric DNA destabilizes H3 nucleosomes to promote CENP-A deposition during the cell cycle
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 215624
DO 10.1101/215624
A1 Manu Shukla
A1 Tong Pin
A1 Sharon A. White
A1 Puneet P. Singh
A1 Angus M. Reid
A1 Catania Sandra
A1 Alison L. Pidoux
A1 Robin C. Allshire
YR 2017
UL http://biorxiv.org/content/early/2017/11/07/215624.abstract
AB Active centromeres are defined by the presence of nucleosomes containing CENP-A, a histone H3 variant, which alone is sufficient to direct kinetochore assembly. Once assembled at a location CENP-A chromatin and kinetochores are maintained at that location though a positive feedback loop where kinetochore proteins recruited by CENP-A itself promote deposition of new CENP-A following replication. Although CENP-A chromatin itself is a heritable entity, it is normally associated with specific sequences. Intrinsic properties of centromeric DNA may favour the assembly of CENP-A rather than H3 nucleosomes. Here we investigate histone dynamics on centromeric DNA. We show that during S-phase histone H3 is deposited as a placeholder at fission yeast centromeres and is subsequently evicted in G2 when we detect deposition of the majority of new CENP-ACnp1. We also find that centromeric DNA has an innate property of driving high rates of turnover of H3 containing nucleosomes resulting in low nucleosome occupancy. When placed at an ectopic chromosomal location in the absence of any CENP-ACnp1 assembly, centromeric DNA retains its ability to impose S-phase deposition and G2 eviction of H3, suggesting that features within this DNA program H3 dynamics. As RNAPII occupancy on this centromere DNA coincides with H3 eviction in G2, we propose a model in which RNAPII-coupled chromatin remodelling promotes replacement of H3 with CENP-ACnp1 nucleosomes.