Conformational landscape of the yeast SAGA complex as revealed by cryo-EM
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s41598-022-16391-0.pdf
Description
Published version
Size
3.93 MB
Format
Adobe PDF
Checksum (MD5)
765d5e16d251425956d84cedd9ae8e27
Author(s)
Vasyliuk, Diana
Felt, Joeseph
Zhong, Ellen D
Berger, Bonnie
Davis, Joseph H
Yip, Calvin K
Date Issued
2022
Journal
Scientific Reports
Publisher
Springer Science and Business Media LLC
Citation
Vasyliuk, Diana, Felt, Joeseph, Zhong, Ellen D, Berger, Bonnie, Davis, Joseph H et al. 2022. "Conformational landscape of the yeast SAGA complex as revealed by cryo-EM." Scientific Reports, 12 (1).
Version
Final published version
Abstract
AbstractSpt-Ada-Gcn5-Acetyltransferase (SAGA) is a conserved multi-subunit complex that activates RNA polymerase II-mediated transcription by acetylating and deubiquitinating nucleosomal histones and by recruiting TATA box binding protein (TBP) to DNA. The prototypical yeast Saccharomyces cerevisiae SAGA contains 19 subunits that are organized into Tra1, core, histone acetyltransferase, and deubiquitination modules. Recent cryo-electron microscopy studies have generated high-resolution structural information on the Tra1 and core modules of yeast SAGA. However, the two catalytical modules were poorly resolved due to conformational flexibility of the full assembly. Furthermore, the high sample requirement created a formidable barrier to further structural investigations of SAGA. Here, we report a workflow for isolating/stabilizing yeast SAGA and preparing cryo-EM specimens at low protein concentration using a graphene oxide support layer. With this procedure, we were able to determine a cryo-EM reconstruction of yeast SAGA at 3.1 Å resolution and examine its conformational landscape with the neural network-based algorithm cryoDRGN. Our analysis revealed that SAGA adopts a range of conformations with its HAT module and central core in different orientations relative to Tra1.
MIT Department
Massachusetts Institute of Technology. Department of Biology
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Creative Commons Attribution 4.0 International license
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DOI of Published Version
https://doi.org/10.1038/S41598-022-16391-0
