Electron microscopy reveals toroidal shape of master neuronal cell differentiator REST - RE1-silencing transcription factor.

电子显微镜显示主神经元细胞分化因子 REST - RE1 沉默转录因子的环形形状。

• 影响指数: 6.155
• 发表时间: 2023
• 来源期刊: Comput Struct Biotechnol J
• DOI: 10.1016/j.csbj.2022.12.026
• PMID: 36698979
• 文章类型: Journal Article
• 中科院分区: Q4
• 方向: 生物
• 作者列表: Veverka P,Brom T,Janovič T,Stojaspal M,Pinkas M,Nováček J,Hofr C

关键词: CD, circular dichroism; CoIP, coimmunoprecipitation; DLS, dynamic light scattering; Differentiation; EM; EM, electron microscopy; Electron microscopy; IDRs, intrinsically disordered regions; NRSE, neuron-restrictive silencer element; NRSF; NRSF, neuron-restrictive silencer factor; Neuron-restrictive silencer factor; Neuronal; PCNA, proliferating cell nuclear antigen; RD1/2, repressor domain 1/2; RE1, repressor element-1; RE1-silencing transcription factor; REST; REST, RE1-silencing transcription factor; REST-FL, full-length REST; REST-N62; REST-N62, splicing isoform of REST, also known as REST4 or REST4-S3; REST4; ZF, zinc finger; aa, amino acid(s); bp, base pair(s); kDa, kilodaltons

Abstract

The RE1-Silencing Transcription factor (REST) is essential for neuronal differentiation. Here, we report the first 18.5-angstrom electron microscopy structure of human REST. The refined electron map suggests that REST forms a torus that can accommodate DNA double-helix in the central hole. Additionally, we quantitatively described REST binding to the canonical DNA sequence of the neuron-restrictive silencer element. We developed protocols for the expression and purification of full-length REST and the shortened variant REST-N62 produced by alternative splicing. We tested the mutual interaction of full-length REST and the splicing variant REST-N62. Revealed structure-function relationships of master neuronal repressor REST will allow finding new biological ways of prevention and treatment of neurodegenerative disorders and diseases.

摘要

RE1沉默转录因子（REST）对于神经元分化至关重要。这里,我们报道了人类REST的第一个18.5埃的电子显微镜结构。精细的电子图表明，REST形成了一个圆环，可以在中心孔中容纳DNA双螺旋。此外,我们定量描述了REST与神经元限制性沉默子元件的规范DNA序列的结合。我们开发了用于表达和纯化全长REST和通过可变剪接产生的缩短变体REST-N62的方案。我们测试了全长REST和剪接变体REST-N62的相互作用。揭示的主神经元阻遏物REST的结构-功能关系将允许寻找预防和治疗神经退行性疾病和疾病的新的生物学方法。