RT Journal Article
SR Electronic
T1 Cohesin depleted cells rebuild functional nuclear compartments after endomitosis
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 816611
DO 10.1101/816611
A1 Marion Cremer
A1 Katharina Brandstetter
A1 Andreas Maiser
A1 Suhas S P Rao
A1 Volker Schmid
A1 Miguel Guirao-Ortiz
A1 Namita Mitra
A1 Stefania Mamberti
A1 Kyle N Klein
A1 David M Gilbert
A1 Heinrich Leonhardt
A1 Maria Cristina Cardoso
A1 Erez Lieberman Aiden
A1 Hartmann Harz
A1 Thomas Cremer
YR 2020
UL http://biorxiv.org/content/early/2020/09/14/816611.abstract
AB Cohesin plays an essential role in chromatin loop extrusion, but its impact on a compartmentalized nuclear architecture, linked to nuclear functions, is debatable. Using live-cell and super-resolved 3D microscopy, we demonstrate that cohesin depleted cells pass through an endomitosis and rebuild a single multilobulated nucleus (MLN) with chromosome territories (CTs) pervaded by interchromatin channels. CTs contain chromatin domain clusters with a zonal organization of repressed chromatin domains in the interior and transcriptionally competent domains located at the periphery. Splicing speckles are located nearby within the lining channel system. These clusters form microscopically defined, active and inactive compartments, which correspond to A/B compartments, detected with ensemble Hi-C. Functionality of MLN despite continuous absence of cohesin was demonstrated by their ability to pass through S-phase with typical spatio-temporal patterns of replication domains. Evidence for structural changes of these domains compared to controls suggests that cohesin is required for their full integrity.Competing Interest StatementThe authors have declared no competing interest.3D FISH3D fluorescence in situ hybridization3D SIM3D structured illumination microscopyAIDauxin inducible degronANC / INCactive / inactive nuclear compartmentCTchromosome territoryCD(C)chromatin domain (cluster)CTCFCCCTC binding factorDAPI4’,6-diamidino-2-phenylindoleEdU5-Ethynyl-2’-deoxyuridineHi-Cchromosome conformation capturing combined with deep sequencingICinterchromatin compartmentMLNmultilobulated nucleusNCnucleosome clusterPBSphosphate buffered salinePBSTphosphate buffered saline with 0.02% TweenPRperichromatin regionRDreplication domainRLreplication labelingTADtopologically associating domain