Cargando…
Transcription factor competition at the γ-globin promoters controls hemoglobin switching
BCL11A, the major regulator of HbF(α(2)γ(2)) level, represses γ-globin expression through direct promoter binding in adult erythroid cells in a switch to adult-type HbA (α(2)β(2)). To uncover how BCL11A initiates repression, we used CRISPR/Cas9, dCas9, dCas9-KRAB, and dCas9-VP64 screens to dissect t...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8038971/ https://www.ncbi.nlm.nih.gov/pubmed/33649594 http://dx.doi.org/10.1038/s41588-021-00798-y |
_version_ | 1783677496358600704 |
---|---|
author | Liu, Nan Xu, Shuqian Yao, Qiuming Zhu, Qian Kai, Yan Hsu, Jonathan Y. Sakon, Phraew Pinello, Luca Yuan, Guo-Cheng Bauer, Daniel E. Orkin, Stuart H. |
author_facet | Liu, Nan Xu, Shuqian Yao, Qiuming Zhu, Qian Kai, Yan Hsu, Jonathan Y. Sakon, Phraew Pinello, Luca Yuan, Guo-Cheng Bauer, Daniel E. Orkin, Stuart H. |
author_sort | Liu, Nan |
collection | PubMed |
description | BCL11A, the major regulator of HbF(α(2)γ(2)) level, represses γ-globin expression through direct promoter binding in adult erythroid cells in a switch to adult-type HbA (α(2)β(2)). To uncover how BCL11A initiates repression, we used CRISPR/Cas9, dCas9, dCas9-KRAB, and dCas9-VP64 screens to dissect the γ-globin promoters and identified an activator element near the BCL11A binding site. Using CUT&RUN and base editing, we demonstrate that a proximal CCAAT box is occupied by the activator NF-Y. BCL11A competes with NF-Y binding through steric hindrance to initiate repression. Occupancy of NF-Y is rapidly established upon BCL11A depletion, and precedes γ-globin derepression and LCR-globin loop formation. Our findings reveal that the switch from fetal-to-adult globin gene expression within the >50 kb β-globin gene cluster is initiated by competition between a stage-selective repressor and a ubiquitous activating factor within a remarkably discrete region of the γ-globin promoters. |
format | Online Article Text |
id | pubmed-8038971 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
record_format | MEDLINE/PubMed |
spelling | pubmed-80389712021-09-01 Transcription factor competition at the γ-globin promoters controls hemoglobin switching Liu, Nan Xu, Shuqian Yao, Qiuming Zhu, Qian Kai, Yan Hsu, Jonathan Y. Sakon, Phraew Pinello, Luca Yuan, Guo-Cheng Bauer, Daniel E. Orkin, Stuart H. Nat Genet Article BCL11A, the major regulator of HbF(α(2)γ(2)) level, represses γ-globin expression through direct promoter binding in adult erythroid cells in a switch to adult-type HbA (α(2)β(2)). To uncover how BCL11A initiates repression, we used CRISPR/Cas9, dCas9, dCas9-KRAB, and dCas9-VP64 screens to dissect the γ-globin promoters and identified an activator element near the BCL11A binding site. Using CUT&RUN and base editing, we demonstrate that a proximal CCAAT box is occupied by the activator NF-Y. BCL11A competes with NF-Y binding through steric hindrance to initiate repression. Occupancy of NF-Y is rapidly established upon BCL11A depletion, and precedes γ-globin derepression and LCR-globin loop formation. Our findings reveal that the switch from fetal-to-adult globin gene expression within the >50 kb β-globin gene cluster is initiated by competition between a stage-selective repressor and a ubiquitous activating factor within a remarkably discrete region of the γ-globin promoters. 2021-03-01 2021-04 /pmc/articles/PMC8038971/ /pubmed/33649594 http://dx.doi.org/10.1038/s41588-021-00798-y Text en http://www.nature.com/authors/editorial_policies/license.html#termsUsers may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms |
spellingShingle | Article Liu, Nan Xu, Shuqian Yao, Qiuming Zhu, Qian Kai, Yan Hsu, Jonathan Y. Sakon, Phraew Pinello, Luca Yuan, Guo-Cheng Bauer, Daniel E. Orkin, Stuart H. Transcription factor competition at the γ-globin promoters controls hemoglobin switching |
title | Transcription factor competition at the γ-globin promoters controls hemoglobin switching |
title_full | Transcription factor competition at the γ-globin promoters controls hemoglobin switching |
title_fullStr | Transcription factor competition at the γ-globin promoters controls hemoglobin switching |
title_full_unstemmed | Transcription factor competition at the γ-globin promoters controls hemoglobin switching |
title_short | Transcription factor competition at the γ-globin promoters controls hemoglobin switching |
title_sort | transcription factor competition at the γ-globin promoters controls hemoglobin switching |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8038971/ https://www.ncbi.nlm.nih.gov/pubmed/33649594 http://dx.doi.org/10.1038/s41588-021-00798-y |
work_keys_str_mv | AT liunan transcriptionfactorcompetitionatthegglobinpromoterscontrolshemoglobinswitching AT xushuqian transcriptionfactorcompetitionatthegglobinpromoterscontrolshemoglobinswitching AT yaoqiuming transcriptionfactorcompetitionatthegglobinpromoterscontrolshemoglobinswitching AT zhuqian transcriptionfactorcompetitionatthegglobinpromoterscontrolshemoglobinswitching AT kaiyan transcriptionfactorcompetitionatthegglobinpromoterscontrolshemoglobinswitching AT hsujonathany transcriptionfactorcompetitionatthegglobinpromoterscontrolshemoglobinswitching AT sakonphraew transcriptionfactorcompetitionatthegglobinpromoterscontrolshemoglobinswitching AT pinelloluca transcriptionfactorcompetitionatthegglobinpromoterscontrolshemoglobinswitching AT yuanguocheng transcriptionfactorcompetitionatthegglobinpromoterscontrolshemoglobinswitching AT bauerdaniele transcriptionfactorcompetitionatthegglobinpromoterscontrolshemoglobinswitching AT orkinstuarth transcriptionfactorcompetitionatthegglobinpromoterscontrolshemoglobinswitching |