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-Structure paper
タイトル | Conformational landscape of the yeast SAGA complex as revealed by cryo-EM. |
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ジャーナル・号・ページ | Sci Rep, Vol. 12, Issue 1, Page 12306, Year 2022 |
掲載日 | 2022年7月19日 |
著者 | Diana Vasyliuk / Joeseph Felt / Ellen D Zhong / Bonnie Berger / Joseph H Davis / Calvin K Yip / |
PubMed 要旨 | Spt-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 ...Spt-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. |
リンク | Sci Rep / PubMed:35853968 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 2.9 - 3.2 Å |
構造データ | EMDB-26792: Transcription co-activator SAGA EMDB-26803: Cryo-EM structure of the SAGA Tra1 module EMDB-26804: Cryo-EM structure of the SAGA core module |
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