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

Authors: Sher, F.; Hossain, M.; Seruggia, D.; Schoonenberg, V. A. C.; Yao, Q.; Cifani, P.; Dassama, L. M. K.; Cole, M. A.; Ren, C.; Vinjamur, D. S.; Macias-Trevino, C.; Luk, K.; McGuckin, C.; Schupp, P. G.; Canver, M. C.; Kurita, R.; Nakamura, Y.; Fujiwara, Y.; Wolfe, S. A.; Pinello, L.; Maeda, T.; Kentsis, A.; Orkin, S. H.; Bauer, D. E.
Article Title: Rational targeting of a NuRD subcomplex guided by comprehensive in situ mutagenesis
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. © 2019, The Author(s), under exclusive licence to Springer Nature America, Inc.
Keywords: genetics; protein domain; mouse; animal; cytology; metabolism; animals; mice; transgenic mouse; mice, transgenic; gene expression regulation; chromatin; erythroid cell; mutagenesis; protein interaction domains and motifs; histone deacetylase; erythroid cells; hemoglobin f; humans; human; fetal hemoglobin; chd4 protein, human; mi-2 nucleosome remodeling and deacetylase complex
Journal Title: Nature Genetics
Volume: 51
Issue: 7
ISSN: 1061-4036
Publisher: Nature Publishing Group  
Date Published: 2019-07-01
Start Page: 1149
End Page: 1159
Language: English
DOI: 10.1038/s41588-019-0453-4
PUBMED: 31253978
PROVIDER: scopus
PMCID: PMC6650275
Notes: Article -- Source: Scopus
Citation Impact
MSK Authors
  1. Alex   Kentsis
    68 Kentsis
  2. Paolo   Cifani
    23 Cifani