@article{abdf93be1bc746df8cea0eac56b4975f,
title = "Direct Promoter Repression by BCL11A Controls the Fetal to Adult Hemoglobin Switch",
abstract = "Fetal hemoglobin (HbF, α2γ2) level is genetically controlled and modifies severity of adult hemoglobin (HbA, α2β2) disorders, sickle cell disease, and β-thalassemia. Common genetic variation affects expression of BCL11A, a regulator of HbF silencing. To uncover how BCL11A supports the developmental switch from γ- to β- globin, we use a functional assay and protein binding microarray to establish a requirement for a zinc-finger cluster in BCL11A in repression and identify a preferred DNA recognition sequence. This motif appears in embryonic and fetal-expressed globin promoters and is duplicated in γ-globin promoters. The more distal of the duplicated motifs is mutated in individuals with hereditary persistence of HbF. Using the CUT&RUN approach to map protein binding sites in erythroid cells, we demonstrate BCL11A occupancy preferentially at the distal motif, which can be disrupted by editing the promoter. Our findings reveal that direct γ-globin gene promoter repression by BCL11A underlies hemoglobin switching. The developmental transition between fetal and adult hemoglobin is controlled by a repressor that acts directly at the γ-globin gene promoter, suggesting a simplified control mechanism that could be manipulated in treatment of γ-hemoglobin disorders.",
keywords = "BCL11A, CUT&RUN, DNA binding, digital genomic footprinting, gene editing, hemoglobin, protein-binding microarray, repression, zinc finger",
author = "Nan Liu and Hargreaves, {Victoria V.} and Qian Zhu and Kurland, {Jesse V.} and Jiyoung Hong and Woojin Kim and Falak Sher and Claudio Macias-Trevino and Rogers, {Julia M.} and Ryo Kurita and Yukio Nakamura and Yuan, {Guo Cheng} and Bauer, {Daniel E.} and Jian Xu and Bulyk, {Martha L.} and Orkin, {Stuart H.}",
note = "Funding Information: We thank Peter Skene and Steven Henikoff for advice on protocols for CUT&RUN, Birgit Knoechel for assistance with DNA sequencing, and Paul Bruno for improvements in protein purification. We appreciate the generosity of Merlin Crossley for sharing findings during the evolution of these studies. Fluorescence polarization experiments were performed at the Institute for Chemistry and Cell Biology-Longwood Screening Facility at HMS. Octet experiments were performed at the Center for Macromolecular Interactions in the Department of Biological Chemistry and Molecular Pharmacology at HMS. The work was supported by the Howard Hughes Medical Institute (HHMI to S.H.O.); National Heart, Lung, and Blood Institute (NHLBI) ( R01 HL119099 to G.C.Y.; R01 HL032259 to S.H.O.); National Human Genome Research Institute (NHGR) ( HG003985I to M.L.B.); National Institute of Diabetes and Digestive and Kidney Disease (NIDDK) ( K01 DK093543 to J.X.; R03 DK109232 to D.E.B.); a Doris Duke Charitable Foundation Innovations in Clinical Research Award to D.E.B. and S.H.O.; and a Cooley's Anemia Foundation Fellowship to D.E.B. Publisher Copyright: {\textcopyright} 2018 Elsevier Inc.",
year = "2018",
month = apr,
day = "5",
doi = "10.1016/j.cell.2018.03.016",
language = "English (US)",
volume = "173",
pages = "430--442.e17",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "2",
}