title: CNOT3-Dependent mRNA Deadenylation Safeguards the Pluripotent State.
abstract: Poly(A) tail length and mRNA deadenylation play important roles in gene regulation. However, how they regulate embryonic development and pluripotent cell fate is not fully understood. Here we present evidence that CNOT3-dependent mRNA deadenylation governs the pluripotent state. We show that CNOT3, a component of the Ccr4-Not deadenylase complex, is required for mouse epiblast maintenance. It is highly expressed in blastocysts and its deletion leads to peri-implantation lethality. The epiblast cells in Cnot3 deletion embryos are quickly lost during diapause and fail to outgrow in culture. Mechanistically, CNOT3 C terminus is required for its interaction with the complex and its function in embryonic stem cells (ESCs). Furthermore, Cnot3 deletion results in increases in the poly(A) tail lengths, half-lives, and steady-state levels of differentiation gene mRNAs. The half-lives of CNOT3 target mRNAs are shorter in ESCs and become longer during normal differentiation. Together, we propose that CNOT3 maintains the pluripotent state by promoting differentiation gene mRNA deadenylation and degradation, and we identify poly(A) tail-length regulation as a post-transcriptional mechanism that controls pluripotency.
authors.affiliation: Epigenetics and Stem Cell Biology Laboratory, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Zheng
authors.affiliation: Epigenetics and Stem Cell Biology Laboratory, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
authors.forename: Pengyi
authors.initials: P
authors.lastname: Yang
authors.affiliation: Epigenetics and Stem Cell Biology Laboratory, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
authors.forename: Brad
authors.initials: B
authors.lastname: Lackford
authors.affiliation: Integrative Bioinformatics Support Group, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Bennett
authors.affiliation: Epigenetics and Stem Cell Biology Laboratory, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.affiliation: Laboratory of Molecular Developmental Biology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: Cellular and Molecular Pathology Branch, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Reproductive and Developmental Biology Laboratory, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
authors.forename: Yiliang
authors.initials: Y
authors.lastname: Miao
authors.affiliation: Cellular and Molecular Pathology Branch, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
authors.forename: Julie F
authors.initials: JF
authors.lastname: Foley
authors.affiliation: Integrative Bioinformatics Support Group, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
authors.forename: David C
authors.initials: DC
authors.lastname: Fargo
authors.affiliation: Laboratory of Molecular Developmental Biology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Reproductive and Developmental Biology Laboratory, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
authors.forename: Carmen J
authors.initials: CJ
authors.lastname: Williams
authors.affiliation: Epigenetics and Stem Cell Biology Laboratory, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
authors.forename: Raja
authors.initials: R
authors.lastname: Jothi
authors.affiliation: Epigenetics and Stem Cell Biology Laboratory, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA. Electronic address: hug4@niehs.nih.gov.
authors.forename: Guang
authors.initials: G
authors.lastname: Hu
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D000066673:Cell Self Renewal / Q000235:genetics; D047108:Embryonic Development / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D017353:Gene Deletion; D018507:Gene Expression Regulation, Developmental; D005855:Germ Layers / Q000196:embryology / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D000072417:Protein Domains / Q000235:genetics; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: C569362:CNOT3 protein, mouse; D012333:RNA, Messenger; D014157:Transcription Factors
keywords: embryonic stem cell; mRNA deadenylation; pluripotent state; pre-implantation development