Absolute quantification of cohesin, CTCF and their regulators in human cells
Year: 2019
Type: article
Abstract: The organisation of mammalian genomes into loops and topologically associating domains (TADs) contributes to chromatin structure, gene expression and recombination. TADs and many loops are formed by cohesin and positioned by CTCF. In proliferating cells, cohesin also mediates sister chromatid cohesion, which is essential for chromosome segregation. Current models of chromatin folding and cohesion are based on assumptions of how many cohesin and CTCF molecules organise the genome. Here we have measured absolute copy numbers and dynamics of cohesin, CTCF, NIPBL, WAPL and sororin by mass spectrometry, fluorescence-correlation spectroscopy and fluorescence recovery after photobleaching in HeLa cells. In G1-phase, there are ~250,000 nuclear cohesin complexes, of which ~ 160,000 are chromatin-bound. Comparison with chromatin immunoprecipitation-sequencing data implies that some genomic cohesin and CTCF enrichment sites are unoccupied in single cells at any one time. We discuss the implications of these findings for how cohesin can contribute to genome organisation and cohesion.
Source: eLife
DOI: 10.7554/elife.46269
Institutions: Institute of Molecular Biotechnology, Royal Netherlands Academy of Arts and Sciences, Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Research Institute of Molecular Pathology, University Medical Center Utrecht, Hubrecht Institute for Developmental Biology and Stem Cell Research, Vienna Biocenter, Novartis (Austria), Max Planck Institute for Biophysical Chemistry, European Molecular Biology Laboratory, European Molecular Biology Laboratory, University of California, Berkeley, Max Planck Institute for Medical Research, University of Gothenburg, Stanford University, University of Oxford, Medical University of Vienna
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,500
Funders: Boehringer Ingelheim, European Molecular Biology Organization, European Molecular Biology Laboratory, Paul G. Allen Frontiers Group, European Commission, Human Frontier Science Program, Vienna Science and Technology Fund, Austrian Science Fund, Österreichische Forschungsförderungsgesellschaft, National Institutes of Health, Horizon 2020 Framework Programme, Biotechnology and Biological Sciences Research Council, Seventh Framework Programme, Sixth Framework Programme, National Institute on Drug Abuse, National Institute of Biomedical Imaging and Bioengineering, Common Fund
Awards: G1334679468, G1639435289, G177484282, Transition of DNA double-strand breaks to genetic crossovers, European Proteomics Infrastructure Consortium providing Access, G2325242462, G2684986193, G2973790809, G3105035837, Chromosome segregation during human cell division, G3207356259, G3441419088, G3583275715, Reconstructing the dynamic 3D architecture of the human genome by superresolution microscopy and DNA sequence modelling., G393177772, Infrastructure for NMR, EM and X-rays for translational research, G4329597992, G4342997908, G434337789, G4626243480, G4875136982, G4937468798, G5255965479, G5702163051, Systems-level characterization of mammalian cell cycle transitions, G601756330, G6341622930, G645210558, G6657544038, G6825305634, Reconstructing the dynamic 3D architecture of the human genome by superresolution microscopy and DNA sequence modelling., G706637122, G7980353291, G8115052865, G8170311165, G823774000, G8243104134, G8260381131, Systems Biology of Mitosis, G8466214923, G8630131099, G8858395165, G983296325
OpenAlex ID: W2949672039