Dataset Information

Temporal resolution of gene derepression and proteome changes upon PROTAC-mediated degradation of BCL11A protein in erythroid cells

ABSTRACT: Reactivation of fetal hemoglobin expression by down-regulation of BCL11A is a promising treatment of -hemoglobinopathies. A detailed understanding of BCL11A-mediated repression of -globin gene (HBG1/2) transcription is lacking, as studies to date used perturbations by shRNA or CRISPR/Cas9 gene editing. We leveraged the dTAG PROTAC platform to acutely deplete BCL11A protein in erythroid cells and examined consequences by PRO-seq, proteomics, chromatin accessibility, and histone profiling. Among ≤ 31 genes repressed by BCL11A, HBG1/2 and HBZ show the most abundant and progressive changes in transcription and chromatin accessibility upon BCL11A loss. Transcriptional changes at HBG1/2 were detected in < 2h. Robust HBG1/2 reactivation upon acute BCL11A-depletion occurred without loss of promoter 5methylcytosine (5mC). Using targeted protein degradation, we establish a hierarchy of gene reactivation at BCL11A targets, in which nascent transcription is followed by increased chromatin accessibility, and both are uncoupled from promoter DNA methylation at the HBG1/2 loci.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Erythrocyte

SUBMITTER: Mario Oroshi

LAB HEAD: Matthias Mann

PROVIDER: PXD030307 | Pride | 2023-10-24

REPOSITORIES: Pride

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			Action	DRS
	20200831_EXPL1_OzKa_SA_Stuti_HUDEP_proteome_noF_LC02_10.raw	Raw
	20200831_EXPL1_OzKa_SA_Stuti_HUDEP_proteome_noF_LC02_11.raw	Raw
	20200831_EXPL1_OzKa_SA_Stuti_HUDEP_proteome_noF_LC02_12.raw	Raw
	20200831_EXPL1_OzKa_SA_Stuti_HUDEP_proteome_noF_LC02_13.raw	Raw
	20200831_EXPL1_OzKa_SA_Stuti_HUDEP_proteome_noF_LC02_14.raw	Raw

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Temporal resolution of gene derepression and proteome changes upon PROTAC-mediated degradation of BCL11A protein in erythroid cells.

Mehta Stuti S Buyanbat Altantsetseg A Kai Yan Y Karayel Ozge O Goldman Seth Raphael SR Seruggia Davide D Zhang Kevin K Fujiwara Yuko Y Donovan Katherine A KA Zhu Qian Q Yang Huan H Nabet Behnam B Gray Nathanael S NS Mann Matthias M Fischer Eric S ES Adelman Karen K Orkin Stuart H SH

Cell chemical biology 20220714 8

Reactivation of fetal hemoglobin expression by the downregulation of BCL11A is a promising treatment for β-hemoglobinopathies. A detailed understanding of BCL11A-mediated repression of γ-globin gene (HBG1/2) transcription is lacking, as studies to date used perturbations by shRNA or CRISPR-Cas9 gene editing. We leveraged the dTAG PROTAC degradation platform to acutely deplete BCL11A protein in erythroid cells and examined consequences by nascent transcriptomics, proteomics, chromatin accessibili ...[more]

PMID: 35839780

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