title: Gene Resistance to Transcriptional Reprogramming following Nuclear Transfer Is Directly Mediated by Multiple Chromatin-Repressive Pathways.
abstract: Understanding the mechanism of resistance of genes to reactivation will help improve the success of nuclear reprogramming. Using mouse embryonic fibroblast nuclei with normal or reduced DNA methylation in combination with chromatin modifiers able to erase H3K9me3, H3K27me3, and H2AK119ub1 from transplanted nuclei, we reveal the basis for resistance of genes to transcriptional reprogramming by oocyte factors. A majority of genes is affected by more than one type of treatment, suggesting that resistance can require repression through multiple epigenetic mechanisms. We classify resistant genes according to their sensitivity to 11 chromatin modifier combinations, revealing the existence of synergistic as well as adverse effects of chromatin modifiers on removal of resistance. We further demonstrate that the chromatin modifier USP21 reduces resistance through its H2AK119 deubiquitylation activity. Finally, we provide evidence that H2A ubiquitylation also contributes to resistance to transcriptional reprogramming in mouse nuclear transfer embryos.
authors.affiliation: Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK. Electronic address: j.jullien@gurdon.cam.ac.uk.
authors.forename: Jerome
authors.initials: J
authors.lastname: Jullien
authors.affiliation: Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK.
authors.forename: Munender
authors.initials: M
authors.lastname: Vodnala
authors.affiliation: Department of Development and Regeneration, KU Leuven, University of Leuven, Herestraat 49, 3000 Leuven, Belgium.
authors.forename: Vincent
authors.initials: V
authors.lastname: Pasque
authors.affiliation: Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK.
authors.forename: Mami
authors.initials: M
authors.lastname: Oikawa
authors.affiliation: Laboratory of Molecular Developmental Biology, Graduate School of Biology-Oriented Science and Technology, Kinki University, Wakayama 649-6493, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Miyamoto
authors.affiliation: Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK.
authors.forename: George
authors.initials: G
authors.lastname: Allen
authors.affiliation: UMR BDR, INRA, ENVA, Université Paris Saclay, 78350 Jouy en Josas, France.
authors.forename: Sarah Anne
authors.initials: SA
authors.lastname: David
authors.affiliation: UMR BDR, INRA, ENVA, Université Paris Saclay, 78350 Jouy en Josas, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Brochard
authors.affiliation: Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK; Laboratory of Muscle Stem Cells and Gene Regulation, National Institute of Arthritis, Musculoskeletal and Skin Diseases (NIAMS), NIH, Bethesda, MD 20892, USA.
authors.forename: Stan
authors.initials: S
authors.lastname: Wang
authors.affiliation: Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK.
authors.forename: Charles
authors.initials: C
authors.lastname: Bradshaw
authors.affiliation: RIKEN Center for Integrative Medical Sciences, Laboratory for Developmental Genetics, North Research Building, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama City, Kanagawa 230-0045, Japan.
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Koseki
authors.affiliation: Laboratory of Muscle Stem Cells and Gene Regulation, National Institute of Arthritis, Musculoskeletal and Skin Diseases (NIAMS), NIH, Bethesda, MD 20892, USA.
authors.forename: Vittorio
authors.initials: V
authors.lastname: Sartorelli
authors.affiliation: UMR BDR, INRA, ENVA, Université Paris Saclay, 78350 Jouy en Josas, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Beaujean
authors.affiliation: Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK.
authors.forename: John
authors.initials: J
authors.lastname: Gurdon
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D065150:Cellular Reprogramming; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly; D003001:Cloning, Molecular; D019175:DNA Methylation; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D044127:Epigenesis, Genetic; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D006657:Histones / Q000378:metabolism*; D008297:Male; D051379:Mice; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D053652:Nuclear Transfer Techniques; D009865:Oocytes; D014158:Transcription, Genetic; D043222:Ubiquitin Thiolesterase / Q000235:genetics / Q000378:metabolism; D054875:Ubiquitination; D014982:Xenopus laevis
chemical_list: D002843:Chromatin; D006657:Histones; C584021:Usp16 protein, mouse; D043222:Ubiquitin Thiolesterase
keywords: chromatin; epigenetic; nuclear transfer; oocyte; resistance; transcriptional reprogramming; xenopus