Rational targeting of a NuRD subcomplex guided by comprehensive in situ mutagenesis

Falak Sher, Mir Hossain, Davide Seruggia, Vivien A.C. Schoonenberg, Qiuming Yao, Paolo Cifani, Laura M.K. Dassama, Mitchel A. Cole, Chunyan Ren, Divya S. Vinjamur, Claudio Macias-Trevino, Kevin Luk, Connor McGuckin, Patrick G. Schupp, Matthew C. Canver, Ryo Kurita, Yukio Nakamura, Yuko Fujiwara, Scot A. Wolfe, Luca PinelloTakahiro Maeda, Alex Kentsis, Stuart H. Orkin, Daniel E. Bauer

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

Developmental silencing of fetal globins serves as both a paradigm of spatiotemporal gene regulation and an opportunity for therapeutic intervention of β-hemoglobinopathy. The nucleosome remodeling and deacetylase (NuRD) chromatin complex participates in γ-globin repression. We used pooled CRISPR screening to disrupt NuRD protein coding sequences comprehensively in human adult erythroid precursors. Essential for fetal hemoglobin (HbF) control is a non-redundant subcomplex of NuRD protein family paralogs, whose composition we corroborated by affinity chromatography and proximity labeling mass spectrometry proteomics. Mapping top functional guide RNAs identified key protein interfaces where in-frame alleles resulted in loss-of-function due to destabilization or altered function of subunits. We ascertained mutations of CHD4 that dissociate its requirement for cell fitness from HbF repression in both primary human erythroid precursors and transgenic mice. Finally we demonstrated that sequestering CHD4 from NuRD phenocopied these mutations. These results indicate a generalizable approach to discover protein complex features amenable to rational biochemical targeting.

Original languageEnglish
Pages (from-to)1149-1159
Number of pages11
JournalNature genetics
Volume51
Issue number7
DOIs
Publication statusPublished - Jul 1 2019

All Science Journal Classification (ASJC) codes

  • Genetics

Fingerprint Dive into the research topics of 'Rational targeting of a NuRD subcomplex guided by comprehensive in situ mutagenesis'. Together they form a unique fingerprint.

Cite this