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SARS-CoV-2 nsp3 and nsp4 are minimal constituents of a pore spanning replication organelle

Author

Listed:
  • Liv Zimmermann

    (Virology, Heidelberg University)

  • Xiaohan Zhao

    (Virology, Heidelberg University)

  • Jana Makroczyova

    (Virology, Heidelberg University)

  • Moritz Wachsmuth-Melm

    (Virology, Heidelberg University)

  • Vibhu Prasad

    (Heidelberg University)

  • Zach Hensel

    (Universidade NOVA de Lisboa)

  • Ralf Bartenschlager

    (Heidelberg University
    German Cancer Research Center (DKFZ)
    Heidelberg partner site)

  • Petr Chlanda

    (Virology, Heidelberg University)

Abstract

Coronavirus replication is associated with the remodeling of cellular membranes, resulting in the formation of double-membrane vesicles (DMVs). A DMV-spanning pore was identified as a putative portal for viral RNA. However, the exact components and the structure of the SARS-CoV-2 DMV pore remain to be determined. Here, we investigate the structure of the DMV pore by in situ cryo-electron tomography combined with subtomogram averaging. We identify non-structural protein (nsp) 3 and 4 as minimal components required for the formation of a DMV-spanning pore, which is dependent on nsp3-4 proteolytic cleavage. In addition, we show that Mac2-Mac3-DPUP-Ubl2 domains are critical for nsp3 oligomerization and crown integrity which influences membrane curvature required for biogenesis of DMVs. Altogether, SARS-CoV-2 nsp3-4 have a dual role by driving the biogenesis of replication organelles and assembly of DMV-spanning pores which we propose here to term replicopores.

Suggested Citation

  • Liv Zimmermann & Xiaohan Zhao & Jana Makroczyova & Moritz Wachsmuth-Melm & Vibhu Prasad & Zach Hensel & Ralf Bartenschlager & Petr Chlanda, 2023. "SARS-CoV-2 nsp3 and nsp4 are minimal constituents of a pore spanning replication organelle," Nature Communications, Nature, vol. 14(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-43666-5
    DOI: 10.1038/s41467-023-43666-5
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