title: MBNL proteins repress ES-cell-specific alternative splicing and reprogramming.
abstract: Previous investigations of the core gene regulatory circuitry that controls the pluripotency of embryonic stem (ES) cells have largely focused on the roles of transcription, chromatin and non-coding RNA regulators. Alternative splicing represents a widely acting mode of gene regulation, yet its role in regulating ES-cell pluripotency and differentiation is poorly understood. Here we identify the muscleblind-like RNA binding proteins, MBNL1 and MBNL2, as conserved and direct negative regulators of a large program of cassette exon alternative splicing events that are differentially regulated between ES cells and other cell types. Knockdown of MBNL proteins in differentiated cells causes switching to an ES-cell-like alternative splicing pattern for approximately half of these events, whereas overexpression of MBNL proteins in ES cells promotes differentiated-cell-like alternative splicing patterns. Among the MBNL-regulated events is an ES-cell-specific alternative splicing switch in the forkhead family transcription factor FOXP1 that controls pluripotency. Consistent with a central and negative regulatory role for MBNL proteins in pluripotency, their knockdown significantly enhances the expression of key pluripotency genes and the formation of induced pluripotent stem cells during somatic cell reprogramming.
authors.affiliation: Banting and Best Department of Medical Research and Donnelly Centre, University of Toronto, Toronto, Ontario M5S 3E1, Canada.
authors.forename: Hong
authors.initials: H
authors.lastname: Han
authors.forename: Manuel
authors.initials: M
authors.lastname: Irimia
authors.forename: P Joel
authors.initials: PJ
authors.lastname: Ross
authors.forename: Hoon-Ki
authors.initials: HK
authors.lastname: Sung
authors.forename: Babak
authors.initials: B
authors.lastname: Alipanahi
authors.forename: Laurent
authors.initials: L
authors.lastname: David
authors.forename: Azadeh
authors.initials: A
authors.lastname: Golipour
authors.forename: Mathieu
authors.initials: M
authors.lastname: Gabut
authors.forename: Iacovos P
authors.initials: IP
authors.lastname: Michael
authors.forename: Emil N
authors.initials: EN
authors.lastname: Nachman
authors.forename: Eric
authors.initials: E
authors.lastname: Wang
authors.forename: Dan
authors.initials: D
authors.lastname: Trcka
authors.forename: Tadeo
authors.initials: T
authors.lastname: Thompson
authors.forename: Dave
authors.initials: D
authors.lastname: O'Hanlon
authors.forename: Valentina
authors.initials: V
authors.lastname: Slobodeniuc
authors.forename: Nuno L
authors.initials: NL
authors.lastname: Barbosa-Morais
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Burge
authors.forename: Jason
authors.initials: J
authors.lastname: Moffat
authors.forename: Brendan J
authors.initials: BJ
authors.lastname: Frey
authors.forename: Andras
authors.initials: A
authors.lastname: Nagy
authors.forename: James
authors.initials: J
authors.lastname: Ellis
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Wrana
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Blencowe
mesh_terms: D017398:Alternative Splicing / Q000235:genetics; D020816:Amino Acid Motifs; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D065150:Cellular Reprogramming; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D051858:Forkhead Transcription Factors / Q000378:metabolism; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; C486626:FOXP1 protein, human; D051858:Forkhead Transcription Factors; C493663:MBNL1 protein, human; C499444:MBNL2 protein, human; C479922:Mbnl1 protein, mouse; C529512:Mbnl2 protein, mouse; D016601:RNA-Binding Proteins; D012097:Repressor Proteins