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

Pavel Veverka; Tomáš Brom; Tomáš Janovič; Martin Stojaspal; Matyáš Pinkas; Jiří Nováček; Ctirad Hofr

doi:10.1016/j.csbj.2022.12.026

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

Comput Struct Biotechnol J. 2022 Dec 19:21:731-741. doi: 10.1016/j.csbj.2022.12.026. eCollection 2023.

Authors

Pavel Veverka^{1

2}, Tomáš Brom¹, Tomáš Janovič¹, Martin Stojaspal^{1

2}, Matyáš Pinkas³, Jiří Nováček³, Ctirad Hofr¹

Affiliations

¹ LifeB, FGP - NCBR, Faculty of Science, Masaryk University, Kamenice 753/5, Brno 625 00, Czech Republic.
² Institute of Biophysics of the Czech Academy of Sciences, Scientific Incubator, Královopolská 135, Brno 612 65, Czech Republic.
³ CEITEC, Masaryk University, Brno, Czech Republic.

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.

Keywords: 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.