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8TXO

E. coli DNA-directed RNA polymerase transcription elongation complex bound to the unnatural dZ-PTP base pair in the active site

8TXO の概要
エントリーDOI10.2210/pdb8txo/pdb
EMDBエントリー41695
分子名称DNA-directed RNA polymerase subunit alpha, DNA-directed RNA polymerase subunit beta, DNA-directed RNA polymerase subunit beta', ... (9 entities in total)
機能のキーワードcomplex, unnatural base pairs, synthetic biology, transcription
由来する生物種Escherichia coli
詳細
タンパク質・核酸の鎖数7
化学式量合計397457.61
構造登録者
Aiyer, S.,Shan, Z.,Oh, J.,Wang, D.,Tan, Y.Z.,Lyumkis, D. (登録日: 2023-08-23, 公開日: 2024-01-24, 最終更新日: 2024-10-30)
主引用文献Aiyer, S.,Baldwin, P.R.,Tan, S.M.,Shan, Z.,Oh, J.,Mehrani, A.,Bowman, M.E.,Louie, G.,Passos, D.O.,Dordevic-Marquardt, S.,Mietzsch, M.,Hull, J.A.,Hoshika, S.,Barad, B.A.,Grotjahn, D.A.,McKenna, R.,Agbandje-McKenna, M.,Benner, S.A.,Noel, J.A.P.,Wang, D.,Tan, Y.Z.,Lyumkis, D.
Overcoming resolution attenuation during tilted cryo-EM data collection.
Nat Commun, 15:389-389, 2024
Cited by
PubMed Abstract: Structural biology efforts using cryogenic electron microscopy are frequently stifled by specimens adopting "preferred orientations" on grids, leading to anisotropic map resolution and impeding structure determination. Tilting the specimen stage during data collection is a generalizable solution but has historically led to substantial resolution attenuation. Here, we develop updated data collection and image processing workflows and demonstrate, using multiple specimens, that resolution attenuation is negligible or significantly reduced across tilt angles. Reconstructions with and without the stage tilted as high as 60° are virtually indistinguishable. These strategies allowed the reconstruction to 3 Å resolution of a bacterial RNA polymerase with preferred orientation, containing an unnatural nucleotide for studying novel base pair recognition. Furthermore, we present a quantitative framework that allows cryo-EM practitioners to define an optimal tilt angle during data acquisition. These results reinforce the utility of employing stage tilt for data collection and provide quantitative metrics to obtain isotropic maps.
PubMed: 38195598
DOI: 10.1038/s41467-023-44555-7
主引用文献が同じPDBエントリー
実験手法
ELECTRON MICROSCOPY (3.1 Å)
構造検証レポート
Validation report summary of 8txo
検証レポート(詳細版)ダウンロードをダウンロード

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件を2025-01-08に公開中

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