title: The short isoform of NF-YA belongs to the embryonic stem cell transcription factor circuitry.
abstract: Totipotency of embryonic stem cells (ESCs) is controlled at the transcriptional level by a handful of transcription factors (TFs) that promote stemness and prevent differentiation. One of the most enriched DNA elements in promoters and enhancers of genes specifically active in ESCs is the CCAAT box, which is recognized by NF-Y, a trimer with histone-like subunits--NF-YB/NF--YC--and the sequence-specific NF-YA. We show that the levels of the short NF-YA isoform--NF-YAs--is high in mouse ESCs (mESCs) and drops after differentiation; a dominant negative mutant affects expression of important stem cells genes, directly and indirectly. Protein transfections of TAT-NF-YAs stimulate growth and compensate for withdrawal of leukemia inhibitory factor (LIF) in cell cultures. Bioinformatic analysis identifies NF-Y sites as highly enriched in genomic loci of stem TFs in ESCs. Specifically, 30%-50% of NANOG peaks have NF-Y sites and indeed NF-Y-binding is required for NANOG association to DNA. These data indicate that NF-Y belongs to the restricted circle of TFs that govern mESCs, and, specifically, that NF-YAs is the active isoform in these cells.
authors.affiliation: Dipartimento di Scienze Biomolecolari e Biotecnologie, Università degli Studi di Milano, Milano, Italy.
authors.forename: Diletta
authors.initials: D
authors.lastname: Dolfini
authors.forename: Mario
authors.initials: M
authors.lastname: Minuzzo
authors.forename: Giulio
authors.initials: G
authors.lastname: Pavesi
authors.forename: Roberto
authors.initials: R
authors.lastname: Mantovani
mesh_terms: D000818:Animals; D023081:CCAAT-Binding Factor / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000378:metabolism* / Q000502:physiology; D015870:Gene Expression; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000502:physiology; D051379:Mice; D000071317:Nanog Homeobox Protein; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism
chemical_list: D023081:CCAAT-Binding Factor; D018398:Homeodomain Proteins; C118391:Klf5 protein, mouse; D051741:Kruppel-Like Transcription Factors; D053605:Leukemia Inhibitory Factor; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; C492060:Nfya protein, mouse; D020033:Protein Isoforms