title: Alternative splicing of MBD2 supports self-renewal in human pluripotent stem cells.
abstract: Alternative RNA splicing (AS) regulates proteome diversity, including isoform-specific expression of several pluripotency genes. Here, we integrated global gene expression and proteomic analyses and identified a molecular signature suggesting a central role for AS in maintaining human pluripotent stem cell (hPSC) self-renewal. We demonstrate that the splicing factor SFRS2 is an OCT4 target gene required for pluripotency. SFRS2 regulates AS of the methyl-CpG binding protein MBD2, whose isoforms play opposing roles in maintenance of and reprogramming to pluripotency. Although both MDB2a and MBD2c are enriched at the OCT4 and NANOG promoters, MBD2a preferentially interacts with repressive NuRD chromatin remodeling factors and promotes hPSC differentiation, whereas overexpression of MBD2c enhances reprogramming of fibroblasts to pluripotency. The miR-301 and miR-302 families provide additional regulation by targeting SFRS2 and MDB2a. These data suggest that OCT4, SFRS2, and MBD2 participate in a positive feedback loop, regulating proteome diversity in support of hPSC self-renewal and reprogramming.
authors.affiliation: Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02115, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Department of Pathology, Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Department of Medicine, Division of Pediatric Hematology Oncology, Howard Hughes Medical Institute, Children's Hospital Boston and Dana-Farber Cancer Institute, Boston, MA 02115, USA; A(∗)STAR Institute of Molecular and Cell Biology and Department of Biological Sciences, National University of Singapore, Singapore 138673, Singapore.
authors.forename: Yuin-Han
authors.initials: YH
authors.lastname: Loh
authors.affiliation: Howard Hughes Medical Institute, Department of Biomedical Engineering and Center of Synthetic Biology, Boston University, Boston, MA 02215, USA; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA; Center for Individualized Medicine, Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Hu
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Department of Medicine, Division of Pediatric Hematology Oncology, Howard Hughes Medical Institute, Children's Hospital Boston and Dana-Farber Cancer Institute, Boston, MA 02115, USA.
authors.forename: Marcella
authors.initials: M
authors.lastname: Cesana
authors.affiliation: Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02115, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Blais Proteomics Center, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
authors.forename: Scott B
authors.initials: SB
authors.lastname: Ficarro
authors.affiliation: Blais Proteomics Center, Dana-Farber Cancer Institute, Boston, MA 02115, USA; Bioinformatics Program, Boston University, Boston, MA 02115, USA.
authors.forename: Jignesh R
authors.initials: JR
authors.lastname: Parikh
authors.affiliation: Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158, USA.
authors.forename: Nathan
authors.initials: N
authors.lastname: Salomonis
authors.affiliation: A(∗)STAR Institute of Molecular and Cell Biology and Department of Biological Sciences, National University of Singapore, Singapore 138673, Singapore.
authors.forename: Cheng-Xu Delon
authors.initials: CX
authors.lastname: Toh
authors.affiliation: Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, Buffalo, NY 14260, USA.
authors.forename: Stelios T
authors.initials: ST
authors.lastname: Andreadis
authors.affiliation: Department of Pathology, Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: C John
authors.initials: CJ
authors.lastname: Luckey
authors.affiliation: Howard Hughes Medical Institute, Department of Biomedical Engineering and Center of Synthetic Biology, Boston University, Boston, MA 02215, USA; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA.
authors.forename: James J
authors.initials: JJ
authors.lastname: Collins
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Department of Medicine, Division of Pediatric Hematology Oncology, Howard Hughes Medical Institute, Children's Hospital Boston and Dana-Farber Cancer Institute, Boston, MA 02115, USA. Electronic address: george.daley@childrens.harvard.edu.
authors.forename: George Q
authors.initials: GQ
authors.lastname: Daley
authors.affiliation: Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02115, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Blais Proteomics Center, Dana-Farber Cancer Institute, Boston, MA 02115, USA. Electronic address: jarrod_marto@dfci.harvard.edu.
authors.forename: Jarrod A
authors.initials: JA
authors.lastname: Marto
mesh_terms: D017398:Alternative Splicing / Q000502:physiology*; D002454:Cell Differentiation; D002470:Cell Survival; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D025461:Feedback, Physiological; D005347:Fibroblasts / Q000502:physiology*; D020869:Gene Expression Profiling; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D000071317:Nanog Homeobox Protein; D009687:Nuclear Proteins / Q000378:metabolism*; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000502:physiology*; D011485:Protein Binding; D020033:Protein Isoforms / Q000235:genetics; D040901:Proteomics; D012261:Ribonucleoproteins / Q000378:metabolism*; D000068103:Serine-Arginine Splicing Factors
chemical_list: D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; C115368:MBD2 protein; C550735:MIRN301A microRNA, human; C532169:MIRN302A microRNA, human; D035683:MicroRNAs; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D009687:Nuclear Proteins; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D020033:Protein Isoforms; D012261:Ribonucleoproteins; C073288:SRSF2 protein, human; D000068103:Serine-Arginine Splicing Factors; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex