Chromatin proteins and RNA are associated with DNA during all phases of mitosis.
Mitosis brings about major changes to chromosome and nuclear structure. We used recently developed proximity ligation assay-based techniques to investigate the association with DNA of chromatin-associated proteins and RNAs inDrosophilaembryos during mitosis. All groups of tested proteins, histone-modifying and chromatin-remodeling proteins and methylated histones remained in close proximity to DNA during all phases of mitosis. We also found that RNA transcripts are associated with DNA during all stages of mitosis. Reduction of H3K27me3 levels or elimination of RNAs had no effect on the association of the components of PcG and TrxG complexes to DNA. Using a combination of proximity ligation assay-based techniques and super-resolution microscopy, we found that the number of protein-DNA and RNA-DNA foci undergoes significant reduction during mitosis, suggesting that mitosis may be accompanied by structural re-arrangement or compaction of specific chromatin domains.
Department of Biochemistry and Molecular Biology and Kimmel Cancer Center, Thomas Jefferson University , Philadelphia, PA, USA.
Kathryn
K
L Black
Department of Biochemistry and Molecular Biology and Kimmel Cancer Center, Thomas Jefferson University , Philadelphia, PA, USA.
Svetlana
S
Petruk
Department of Biochemistry and Molecular Biology and Kimmel Cancer Center, Thomas Jefferson University , Philadelphia, PA, USA.
Tyler K
TK
Fenstermaker
Department of Zoology, University of British Columbia , Vancouver, BC, Canada.
Jacob W
JW
Hodgson
Delaware Biotechnology Institute Bio-Imaging Center, University of Delaware , Newark, DE, USA.
Jeffrey L
JL
Caplan
Department of Zoology, University of British Columbia , Vancouver, BC, Canada.
Hugh W
HW
Brock
Department of Biochemistry and Molecular Biology and Kimmel Cancer Center, Thomas Jefferson University , Philadelphia, PA, USA.
Alexander
A
Mazo
PcG; TrxG; chromatin; epigenetics; mitosis