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 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 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 <2 h. Robust HBG1/2 reactivation upon acute BCL11A depletion occurred without the loss of promoter 5-methylcytosine (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.

• Authors: Stuti Mehta, Altantsetseg Buyanbat, Yan Kai, Ozge Karayel, Seth Raphael Goldman, Davide Seruggia, Kevin Zhang, Yuko Fujiwara, Katherine A Donovan, Qian Zhu, Huan Yang, Behnam Nabet, Nathanael S Gray, Matthias Mann, Eric S Fischer, Karen Adelman, Stuart H Orkin
• Journal: Cell chemical biology (Cell Chem Biol) Vol. 29 Issue 8 Pg. 1273-1287.e8 (08 18 2022) ISSN: 2451-9448 [Electronic] United States
• PMID: 35839780 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.

Chemical References

• Carrier Proteins
• Chromatin
• Nuclear Proteins
• Proteome
• Repressor Proteins
• Transcription Factors

Topics

• Carrier Proteins (metabolism)
• Chromatin (genetics, metabolism)
• Erythroid Cells (metabolism)
• Nuclear Proteins (metabolism)
• Proteome (metabolism)
• Repressor Proteins (genetics, metabolism)
• Transcription Factors (metabolism)