哈佛医学院Daniel E. Bauer研究团队提出了在综合原位诱变的指导下合理定位NuRD亚复合物。 相关论文于2019年7月发表于国际学术期刊《自然—遗传学》上。

研究组使用集合CRISPR筛选来全面破坏成人红细胞前体中的NuRD蛋白编码序列。胎儿血红蛋白（HbF）控制的必要条件是NuRD蛋白家族旁系同源物的非冗余亚复合物，研究小组通过亲和层析和邻近标记质谱蛋白质组学证实了其组成。绘制顶级功能指导RNAs识别关键蛋白质界面，其中框架内等位基因由于亚单位的不稳定或功能改变而导致功能丧失。我们确定了CHD4的突变，其对细胞适应性的要求与原代人红细胞前体和转基因小鼠的HbF抑制无关。最后，我们证明了从NuRD现象中隔离CHD4复制了这些突变。这些结果表明了一种发现适合合理生化靶向的蛋白质复合物特征的通用方法。

据悉，胎儿球蛋白的发育沉默既是时空基因调控的范例，也是β-血红蛋白病治疗干预的机会，核小体重塑和去乙酰化酶(NuRD)染色质复合体参与了γ-珠蛋白的抑制。

附：英文原文

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

Author: 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 Pinello, Takahiro Maeda, Alex Kentsis, Stuart H. Orkin, Daniel E. Bauer

Issue&Volume: Volume 51 Issue 7, July 2019

Abstract: Developmental silencing of fetal globins serves as both a paradigm of spatiotemporal gene regulation and an opportunity for therapeutic intervention of -hemoglobinopathy. The nucleosome remodeling and deacetylase (NuRD) chromatin complex participates in -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.

DOI: 10.1038/s41588-019-0453-4

Source: https://www.nature.com/articles/s41588-019-0453-4