title: RUNX1 regulates the CD34 gene in haematopoietic stem cells by mediating interactions with a distal regulatory element.
abstract: The transcription factor RUNX1 is essential to establish the haematopoietic gene expression programme; however, the mechanism of how it activates transcription of haematopoietic stem cell (HSC) genes is still elusive. Here, we obtained novel insights into RUNX1 function by studying regulation of the human CD34 gene, which is expressed in HSCs. Using transgenic mice carrying human CD34 PAC constructs, we identified a novel downstream regulatory element (DRE), which is bound by RUNX1 and is necessary for human CD34 expression in long-term (LT)-HSCs. Conditional deletion of Runx1 in mice harbouring human CD34 promoter-DRE constructs abrogates human CD34 expression. We demonstrate by chromosome conformation capture assays in LT-HSCs that the DRE physically interacts with the human CD34 promoter. Targeted mutagenesis of RUNX binding sites leads to perturbation of this interaction and decreased human CD34 expression in LT-HSCs. Overall, our in vivo data provide novel evidence about the role of RUNX1 in mediating interactions between distal and proximal elements of the HSC gene CD34.
authors.affiliation: Division of Hematology/Oncology, Beth Israel Deaconess Medical Center, Center for Life Science, Boston, MA, USA. elevanti@bidmc.harvard.edu
authors.forename: Elena
authors.initials: E
authors.lastname: Levantini
authors.forename: Sanghoon
authors.initials: S
authors.lastname: Lee
authors.forename: Hanna S
authors.initials: HS
authors.lastname: Radomska
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Hetherington
authors.forename: Meritxell
authors.initials: M
authors.lastname: Alberich-Jorda
authors.forename: Giovanni
authors.initials: G
authors.lastname: Amabile
authors.forename: Pu
authors.initials: P
authors.lastname: Zhang
authors.forename: David A
authors.initials: DA
authors.lastname: Gonzalez
authors.forename: Junyan
authors.initials: J
authors.lastname: Zhang
authors.forename: Daniela S
authors.initials: DS
authors.lastname: Basseres
authors.forename: Nicola K
authors.initials: NK
authors.lastname: Wilson
authors.forename: Steffen
authors.initials: S
authors.lastname: Koschmieder
authors.forename: Gang
authors.initials: G
authors.lastname: Huang
authors.forename: Dong-Er
authors.initials: DE
authors.lastname: Zhang
authors.forename: Alexander K
authors.initials: AK
authors.lastname: Ebralidze
authors.forename: Constanze
authors.initials: C
authors.lastname: Bonifer
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Okuno
authors.forename: Bertie
authors.initials: B
authors.lastname: Gottgens
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Tenen
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism*; D016026:Bone Marrow Transplantation; D002843:Chromatin / Q000378:metabolism; D050676:Core Binding Factor Alpha 2 Subunit / Q000235:genetics* / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology; D005786:Gene Expression Regulation; D005838:Genotype; D018922:HL-60 Cells; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D008954:Models, Biological; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics
chemical_list: D018952:Antigens, CD34; D002843:Chromatin; D050676:Core Binding Factor Alpha 2 Subunit; C493728:RUNX1 protein, human; C493726:Runx1 protein, mouse