TY - JOUR
T1 - Human centromeric CENP-A chromatin is a homotypic, octameric nucleosome at all cell cycle points
AU - Nechemia-Arbely, Yael
AU - Fachinetti, Daniele
AU - Miga, Karen H.
AU - Sekulic, Nikolina
AU - Soni, Gautam V.
AU - Kim, Dong Hyun
AU - Wong, Adeline K.
AU - Lee, Ah Young
AU - Nguyen, Kristen
AU - Dekker, Cees
AU - Ren, Bing
AU - Black, Ben E.
AU - Cleveland, Don W.
PY - 2017/3/6
Y1 - 2017/3/6
N2 - Chromatin assembled with centromere protein A (CENP-A) is the epigenetic mark of centromere identity. Using new reference models, we now identify sites of CENP-A and histone H3.1 binding within the megabase, α-satellite repeat-containing centromeres of 23 human chromosomes. The overwhelming majority (97%) of α-satellite DNA is found to be assembled with histone H3.1-containing nucleosomes with wrapped DNA termini. In both G1 and G2 cell cycle phases, the 2-4% of α-satellite assembled with CENP-A protects DNA lengths centered on 133 bp, consistent with octameric nucleosomes with DNA unwrapping at entry and exit. CENP-A chromatin is shown to contain equimolar amounts of CENP-A and histones H2A, H2B, and H4, with no H3. Solid-state nanopore analyses show it to be nucleosomal in size. Thus, in contrast to models for hemisomes that briefly transition to octameric nucleosomes at specific cell cycle points or heterotypic nucleosomes containing both CENP-A and histone H3, human CENP-A chromatin complexes are octameric nucleosomes with two molecules of CENP-A at all cell cycle phases.
AB - Chromatin assembled with centromere protein A (CENP-A) is the epigenetic mark of centromere identity. Using new reference models, we now identify sites of CENP-A and histone H3.1 binding within the megabase, α-satellite repeat-containing centromeres of 23 human chromosomes. The overwhelming majority (97%) of α-satellite DNA is found to be assembled with histone H3.1-containing nucleosomes with wrapped DNA termini. In both G1 and G2 cell cycle phases, the 2-4% of α-satellite assembled with CENP-A protects DNA lengths centered on 133 bp, consistent with octameric nucleosomes with DNA unwrapping at entry and exit. CENP-A chromatin is shown to contain equimolar amounts of CENP-A and histones H2A, H2B, and H4, with no H3. Solid-state nanopore analyses show it to be nucleosomal in size. Thus, in contrast to models for hemisomes that briefly transition to octameric nucleosomes at specific cell cycle points or heterotypic nucleosomes containing both CENP-A and histone H3, human CENP-A chromatin complexes are octameric nucleosomes with two molecules of CENP-A at all cell cycle phases.
UR - http://www.scopus.com/inward/record.url?scp=85021847585&partnerID=8YFLogxK
UR - http://resolver.tudelft.nl/uuid:19164e31-0f3e-49d6-b128-3af464f3cfd2
U2 - 10.1083/jcb.201608083
DO - 10.1083/jcb.201608083
M3 - Article
C2 - 28235947
AN - SCOPUS:85021847585
SN - 0021-9525
VL - 216
SP - 607
EP - 621
JO - The Journal of Cell Biology
JF - The Journal of Cell Biology
IS - 3
ER -