Protocol for subtomogram averaging of helical filaments in cryo-electron tomography

被引：1

作者：

Zhang, Xiaojie ^{[1
,3
,4
]}

Mahamid, Julia ^{[1
,2
]}

机构：

[1] European Mol Biol Lab, Struct & Computat Biol Unit, D-69117 Heidelberg, Baden Wurttembe, Germany

[2] European Mol Biol Lab, Cell Biol & Biophys Unit, D-69117 Heidelberg, Baden Wurttembe, Germany

[3] ShanghaiTech Univ, iHuman Inst, Sch Life Sci & Technol, Shanghai 201210, Peoples R China

[4] ShanghaiTech Univ, Shanghai Key Lab High Resolut Electron Microscopy, Shanghai 201210, Peoples R China

来源：

STAR PROTOCOLS | 2024年 / 5卷 / 03期

基金：

欧洲研究理事会;

关键词：

IN-SITU; ELECTRON; VISUALIZATION;

D O I：

10.1016/j.xpro.2024.103272

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Helical filaments are essential macromolecular elements in cellular organization and dynamics. Recent advances in cryo-electron tomography allow faithful imaging of isolated or in-cell filaments. Here, we present a protocol to generate density maps at sub-nanometer resolution of helical filaments by subtomogram averaging, exemplified with isolated mumps virus nucleocapsids and their in- cell form as an extension of the protocol. We detail procedures from pre-processing of tilt-series movie frames to refinement of reconstructed averages for streamlined data processing of helical filaments. For complete details on the use and execution of this protocol, please refer to Zhang et al.1 1

引用

页数：33

共 30 条

[21] Multi-particle cryo-EM refinement with M visualizes ribosome-antibiotic complex at 3.5 Å in cells [J].

Tegunov, Dimitry ;

Xue, Liang ;

Dienemann, Christian ;

Cramer, Patrick ;

Mahamid, Julia .

NATURE METHODS, 2021, 18 (02) :186-+

[22] Real-time cryo-electron microscopy data preprocessing with Warp [J].

Tegunov, Dimitry ;

Cramer, Patrick .

NATURE METHODS, 2019, 16 (11) :1146-+

[23] Structure and assembly of the Ebola virus nucleocapsid [J].

Wan, William ;

Kolesnikova, Larissa ;

Clarke, Mairi ;

Koehler, Alexander ;

Noda, Takeshi ;

Becker, Stephan ;

Briggs, John A. G. .

NATURE, 2017, 551 (7680) :394-+

[24] Structures from intact myofibrils reveal mechanism of thin filament regulation through nebulin [J].

Wang, Zhexin ;

Grange, Michael ;

Pospich, Sabrina ;

Wagner, Thorsten ;

Kho, Ay Lin ;

Gautel, Mathias ;

Raunser, Stefan .

SCIENCE, 2022, 375 (6582) :738-+

[25] The In Situ Structure of Parkinson's Disease-Linked LRRK2 [J].

Watanabe, Reika ;

Buschauer, Robert ;

Bohning, Jan ;

Audagnotto, Martina ;

Lasker, Keren ;

Lu, Tsan-Wen ;

Boassa, Daniela ;

Taylor, Susan ;

Villa, Elizabeth .

CELL, 2020, 182 (06) :1508-+

[26] The structure of the COPII transport-vesicle coat assembled on membranes [J].

Zanetti, Giulia ;

Prinz, Simone ;

Daum, Sebastian ;

Meister, Annette ;

Schekman, Randy ;

Bacia, Kirsten ;

Briggs, John A. G. .

ELIFE, 2013, 2

[27] Molecular mechanisms of stress-induced reactivation in mumps virus condensates [J].

Zhang, Xiaojie ;

Sridhran, Sindhuja ;

Zagoriy, Levgeniia ;

Oegema, Christina Eugster ;

Ching, Cyan ;

Pflaesterer, Tim ;

Fung, Herman K. H. ;

Becher, Isabelle ;

Poser, Ina ;

Mueller, Christoph W. ;

Hyman, Anthony A. ;

Savilski, Mikhil M. ;

Mahamid, Julia .

CELL, 2023, 186 (09) :1877-+

[28] AreTomo: An integrated software package for automated marker-free, motion-corrected cryo-electron tomographic alignment and reconstruction [J].

Zheng, Shawn ;

Wolff, Georg ;

Greenan, Garrett ;

Chen, Zhen ;

Faas, Frank G. A. ;

Barcena, Montserrat ;

Koster, Abraham J. ;

Cheng, Yifan ;

Agard, David A. .

JOURNAL OF STRUCTURAL BIOLOGY-X, 2022, 6

[29] A Bayesian approach to single-particle electron cryo-tomography in RELION-4.0 [J].

Zivanov, Jasenko ;

Oton, Joaquin ;

Ke, Zunlong ;

von Kuegelgen, Andriko ;

Pyle, Euan ;

Qu, Kun ;

Morado, Dustin ;

Castano-Diez, Daniel ;

Zanetti, Giulia ;

Bharat, Tanmay A. M. ;

Briggs, John A. G. ;

Scheres, Sjors H. W. .

ELIFE, 2022, 11

[30] New tools for automated high-resolution cryo-EM structure determination in RELION-3 [J].

Zivanov, Jasenko ;

Nakane, Takanori ;

Forsberg, Bjorn O. ;

Kimanius, Dari ;

Hagen, Wim J. H. ;

Lindahl, Erik ;

Scheres, Sjors H. W. .

ELIFE, 2018, 7

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