Structure of the ciliary axoneme at nanometer resolution reconstructed by TYGRESS

Kangkang Song; Zhiguo Shang; Xiaofeng Fu; Xiaochu Lou; Nikolaus Grigorieff; Daniela Nicastro

doi:10.1101/363317

Structure of the ciliary axoneme at nanometer resolution reconstructed by TYGRESS

Kangkang Song, Zhiguo Shang, Xiaofeng Fu, Xiaochu Lou, Nikolaus Grigorieff, Daniela Nicastro

Research output: Contribution to journal › Article › peer-review

Abstract

The resolution of subtomogram averages calculated from cryo-electron tomograms (cryo-ET) of crowded cellular environments is often limited due to signal loss in, and misalignment of the subtomograms. In contrast, single-particle cryo-electron microcopy (SP-cryo-EM) routinely reaches near-atomic resolution of isolated complexes. We developed a novel hybrid-method called “TomographY-Guided 3D REconstruction of Subcellular Structures” (TYGRESS) that combines cryo-ET with SP-cryo-EM to achieve close-to-nanometer resolution of complexes inside crowded environments. Using TYGRESS, we determined the native 3D structures of the intact ciliary axoneme with up to 12 Å resolution. These results reveal many structures and details that were not visible by cryo-ET. TYGRESS is generally applicable to cellular complexes that are amenable to subtomogram averaging, bringing us a step closer to (pseudo-)atomic models of cells. One Sentence Summary A hybrid cryo-electron microscopy method reveals subcellular structures at unprecedented resolution.

Original language	English (US)
Journal	Unknown Journal
DOIs	https://doi.org/10.1101/363317
State	Published - Jul 6 2018

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Agricultural and Biological Sciences(all)
Immunology and Microbiology(all)
Neuroscience(all)
Pharmacology, Toxicology and Pharmaceutics(all)

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title = "Structure of the ciliary axoneme at nanometer resolution reconstructed by TYGRESS",

abstract = "The resolution of subtomogram averages calculated from cryo-electron tomograms (cryo-ET) of crowded cellular environments is often limited due to signal loss in, and misalignment of the subtomograms. In contrast, single-particle cryo-electron microcopy (SP-cryo-EM) routinely reaches near-atomic resolution of isolated complexes. We developed a novel hybrid-method called “TomographY-Guided 3D REconstruction of Subcellular Structures” (TYGRESS) that combines cryo-ET with SP-cryo-EM to achieve close-to-nanometer resolution of complexes inside crowded environments. Using TYGRESS, we determined the native 3D structures of the intact ciliary axoneme with up to 12 {\AA} resolution. These results reveal many structures and details that were not visible by cryo-ET. TYGRESS is generally applicable to cellular complexes that are amenable to subtomogram averaging, bringing us a step closer to (pseudo-)atomic models of cells. One Sentence Summary A hybrid cryo-electron microscopy method reveals subcellular structures at unprecedented resolution.",

author = "Kangkang Song and Zhiguo Shang and Xiaofeng Fu and Xiaochu Lou and Nikolaus Grigorieff and Daniela Nicastro",

note = "Publisher Copyright: The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

year = "2018",

month = jul,

day = "6",

doi = "10.1101/363317",

language = "English (US)",

journal = "Seminars in Fetal and Neonatal Medicine",

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AU - Song, Kangkang

AU - Shang, Zhiguo

AU - Fu, Xiaofeng

AU - Lou, Xiaochu

AU - Grigorieff, Nikolaus

AU - Nicastro, Daniela

N1 - Publisher Copyright: The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.

PY - 2018/7/6

Y1 - 2018/7/6

N2 - The resolution of subtomogram averages calculated from cryo-electron tomograms (cryo-ET) of crowded cellular environments is often limited due to signal loss in, and misalignment of the subtomograms. In contrast, single-particle cryo-electron microcopy (SP-cryo-EM) routinely reaches near-atomic resolution of isolated complexes. We developed a novel hybrid-method called “TomographY-Guided 3D REconstruction of Subcellular Structures” (TYGRESS) that combines cryo-ET with SP-cryo-EM to achieve close-to-nanometer resolution of complexes inside crowded environments. Using TYGRESS, we determined the native 3D structures of the intact ciliary axoneme with up to 12 Å resolution. These results reveal many structures and details that were not visible by cryo-ET. TYGRESS is generally applicable to cellular complexes that are amenable to subtomogram averaging, bringing us a step closer to (pseudo-)atomic models of cells. One Sentence Summary A hybrid cryo-electron microscopy method reveals subcellular structures at unprecedented resolution.

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