title: Deterministic Restriction on Pluripotent State Dissolution by Cell-Cycle Pathways.
abstract: During differentiation, human embryonic stem cells (hESCs) shut down the regulatory network conferring pluripotency in a process we designated pluripotent state dissolution (PSD). In a high-throughput RNAi screen using an inclusive set of differentiation conditions, we identify centrally important and context-dependent processes regulating PSD in hESCs, including histone acetylation, chromatin remodeling, RNA splicing, and signaling pathways. Strikingly, we detected a strong and specific enrichment of cell-cycle genes involved in DNA replication and G2 phase progression. Genetic and chemical perturbation studies demonstrate that the S and G2 phases attenuate PSD because they possess an intrinsic propensity toward the pluripotent state that is independent of G1 phase. Our data therefore functionally establish that pluripotency control is hardwired to the cell-cycle machinery, where S and G2 phase-specific pathways deterministically restrict PSD, whereas the absence of such pathways in G1 phase potentially permits the initiation of differentiation.
authors.affiliation: Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore; National University of Singapore Graduate School for Integrative Sciences and Engineering, National University of Singapore, 28 Medical Drive, Singapore 117456, Singapore.
authors.forename: Kevin Andrew Uy
authors.initials: KA
authors.lastname: Gonzales
authors.affiliation: Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore.
authors.forename: Hongqing
authors.initials: H
authors.lastname: Liang
authors.affiliation: Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore.
authors.forename: Yee-Siang
authors.initials: YS
authors.lastname: Lim
authors.affiliation: Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore.
authors.forename: Yun-Shen
authors.initials: YS
authors.lastname: Chan
authors.affiliation: Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 639798, Singapore.
authors.forename: Jia-Chi
authors.initials: JC
authors.lastname: Yeo
authors.affiliation: Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore.
authors.forename: Cheng-Peow
authors.initials: CP
authors.lastname: Tan
authors.affiliation: Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore.
authors.forename: Bin
authors.initials: B
authors.lastname: Gao
authors.affiliation: Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore; Department of Biochemistry, National University of Singapore, 8 Medical Drive, Singapore 117597.
authors.forename: Beilin
authors.initials: B
authors.lastname: Le
authors.affiliation: Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore; Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore.
authors.forename: Zi-Ying
authors.initials: ZY
authors.lastname: Tan
authors.affiliation: School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 639798, Singapore.
authors.forename: Kok-Yao
authors.initials: KY
authors.lastname: Low
authors.affiliation: National University of Singapore Graduate School for Integrative Sciences and Engineering, National University of Singapore, 28 Medical Drive, Singapore 117456, Singapore; Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore.
authors.forename: Yih-Cherng
authors.initials: YC
authors.lastname: Liou
authors.affiliation: Department of Biochemistry, National University of Singapore, 8 Medical Drive, Singapore 117597; Institute of Molecular and Cellular Biology, 61 Biopolis Drive, Singapore 138673, Singapore.
authors.forename: Frederic
authors.initials: F
authors.lastname: Bard
authors.affiliation: Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore; Department of Biochemistry, National University of Singapore, 8 Medical Drive, Singapore 117597; Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 639798, Singapore. Electronic address: nghh@gis.a-star.edu.sg.
authors.forename: Huck-Hui
authors.initials: HH
authors.lastname: Ng
mesh_terms: D064007:Ataxia Telangiectasia Mutated Proteins / Q000378:metabolism; D002453:Cell Cycle; D002454:Cell Differentiation; D056765:Cyclin B2 / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D044127:Epigenesis, Genetic; D053263:Gene Regulatory Networks; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: C503758:CCNB2 protein, human; D056765:Cyclin B2; D016159:Tumor Suppressor Protein p53; C576324:ATM protein, human; C576597:ATR protein, human; D064007:Ataxia Telangiectasia Mutated Proteins