title: Noncanonical function of DGCR8 controls mESC exit from pluripotency.
abstract: Mouse embryonic stem cells (mESCs) deficient for DGCR8, a key component of the microprocessor complex, present strong differentiation defects. However, the exact reasons impairing their commitment remain elusive. The analysis of newly generated mutant mESCs revealed that DGCR8 is essential for the exit from the pluripotency state. To dissociate canonical versus noncanonical functions of DGCR8, we complemented the mutant mESCs with a phosphomutant DGCR8, which restored microRNA levels but did not rescue the exit from pluripotency defect. Integration of omics data and RNA immunoprecipitation experiments established DGCR8 as a direct interactor of Tcf7l1 mRNA, a core component of the pluripotency network. Finally, we found that DGCR8 facilitated the splicing of Tcf7l1, an event necessary for the differentiation of mESCs. Our data reveal a new noncanonical function of DGCR8 in the modulation of the alternative splicing of Tcf7l1 mRNA in addition to its established function in microRNA biogenesis.
authors.affiliation: Department of Biology, Institute of Molecular Health Sciences, RNAi and Genome Integrity, Swiss Federal Institute of Technology Zurich, Zurich 8093, Switzerland.
authors.forename: Daniel
authors.identifier: 0000-0003-2171-5032
authors.initials: D
authors.lastname: Cirera-Salinas
authors.affiliation: Department of Biology, Institute of Molecular Health Sciences, RNAi and Genome Integrity, Swiss Federal Institute of Technology Zurich, Zurich 8093, Switzerland.|Life Science Zurich Graduate School, University of Zurich, Zurich 8093, Switzerland.
authors.forename: Jian
authors.initials: J
authors.lastname: Yu
authors.affiliation: Department of Biology, Institute of Molecular Health Sciences, RNAi and Genome Integrity, Swiss Federal Institute of Technology Zurich, Zurich 8093, Switzerland.|Life Science Zurich Graduate School, University of Zurich, Zurich 8093, Switzerland.
authors.forename: Maxime
authors.identifier: 0000-0003-0500-4767
authors.initials: M
authors.lastname: Bodak
authors.affiliation: Department of Biology, Institute of Molecular Health Sciences, RNAi and Genome Integrity, Swiss Federal Institute of Technology Zurich, Zurich 8093, Switzerland.
authors.forename: Richard P
authors.identifier: 0000-0003-4847-9375
authors.initials: RP
authors.lastname: Ngondo
authors.affiliation: Sanford Burnham Prebys Medical Discovery Institute, La Jolla, San Diego, CA 92037.
authors.forename: Kristina M
authors.identifier: 0000-0002-5288-3381
authors.initials: KM
authors.lastname: Herbert
authors.affiliation: Department of Biology, Institute of Molecular Health Sciences, RNAi and Genome Integrity, Swiss Federal Institute of Technology Zurich, Zurich 8093, Switzerland cciaudo@ethz.ch.
authors.forename: Constance
authors.identifier: 0000-0002-0857-4506
authors.initials: C
authors.lastname: Ciaudo
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D002453:Cell Cycle; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D053595:Embryonic Stem Cells / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D005838:Genotype; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics; D009154:Mutation; D010641:Phenotype; D010766:Phosphorylation; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D011485:Protein Binding; D034622:RNA Interference; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013997:Time Factors; D058125:Transcription Factor 7-Like 1 Protein / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: C474044:Dgcr8 protein, mouse; D035683:MicroRNAs; D012322:RNA Precursors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C544913:Tcf7l1 protein, mouse; D058125:Transcription Factor 7-Like 1 Protein