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    A microtubule RELION-based pipeline for cryo-EM image processing

    Cook, A.D. and Manka, Szymon W. and Wang, S. and Moores, Carolyn A. and Atherton, Joe (2019) A microtubule RELION-based pipeline for cryo-EM image processing. Journal of Structural Biology , ISSN 1047-8477. (In Press)

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    Abstract

    Microtubules are polar filaments built from αβ-tubulin heterodimers that exhibit a range of architectures in vitro and in vivo. Tubulin heterodimers are arranged helically in the microtubule wall but many physiologically relevant architectures exhibit a break in helical symmetry known as the seam. Noisy 2D cryo-electron microscopy projection images of pseudo-helical microtubules therefore depict distinct but highly similar views owing to the high structural similarity of α- and β-tubulin. The determination of the αβ-tubulin register and seam location during image processing is essential for alignment accuracy that enables determination of biologically relevant structures. Here we present a pipeline designed for image processing and high-resolution reconstruction of cryo-electron microscopy microtubule datasets, based in the popular and user-friendly RELION image-processing package, Microtubule RELION-based Pipeline (MiRP). The pipeline uses a combination of supervised classification and prior knowledge about geometric lattice constraints in microtubules to accurately determine microtubule architecture and seam location. The presented method is fast and semi-automated, producing near-atomic resolution reconstructions with test datasets that contain a range of microtubule architectures and binding proteins.

    Metadata

    Item Type: Article
    Keyword(s) / Subject(s): Microtubule, pseudo-helical symmetry, cryo-EM, RELION, 3D reconstruction
    School: Birkbeck Schools and Departments > School of Science > Biological Sciences
    Depositing User: Administrator
    Date Deposited: 16 Oct 2019 11:19
    Last Modified: 05 Dec 2019 19:38
    URI: http://eprints.bbk.ac.uk/id/eprint/29525

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