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Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase

Author

Listed:
  • Ning Cui

    (Southern University of Science and Technology)

  • Jun-Tao Zhang

    (Southern University of Science and Technology)

  • Zhuolin Li

    (Southern University of Science and Technology)

  • Xiao-Yu Liu

    (Southern University of Science and Technology)

  • Chongyuan Wang

    (Chinese Academy of Science
    Chinese Academy of Science)

  • Hongda Huang

    (Southern University of Science and Technology)

  • Ning Jia

    (Southern University of Science and Technology
    Guangdong Provincial Key Laboratory of Cell Microenvironment and Disease Research, Southern University of Science and Technology
    Key University Laboratory of Metabolism and Health of Guangdong, Southern University of Science and Technology)

Abstract

The RNA-targeting type III-E CRISPR-gRAMP effector interacts with a caspase-like protease TPR-CHAT to form the CRISPR-guided caspase complex (Craspase), but their functional mechanism is unknown. Here, we report cryo-EM structures of the type III-E gRAMPcrRNA and gRAMPcrRNA-TPR-CHAT complexes, before and after either self or non-self RNA target binding, and elucidate the mechanisms underlying RNA-targeting and non-self RNA-induced protease activation. The associated TPR-CHAT adopted a distinct conformation upon self versus non-self RNA target binding, with nucleotides at positions −1 and −2 of the CRISPR-derived RNA (crRNA) serving as a sensor. Only binding of the non-self RNA target activated the TPR-CHAT protease, leading to cleavage of Csx30 protein. Furthermore, TPR-CHAT structurally resembled eukaryotic separase, but with a distinct mechanism for protease regulation. Our findings should facilitate the development of gRAMP-based RNA manipulation tools, and advance our understanding of the virus-host discrimination process governed by a nuclease-protease Craspase during type III-E CRISPR-Cas immunity.

Suggested Citation

  • Ning Cui & Jun-Tao Zhang & Zhuolin Li & Xiao-Yu Liu & Chongyuan Wang & Hongda Huang & Ning Jia, 2022. "Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
  • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-35275-5
    DOI: 10.1038/s41467-022-35275-5
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    References listed on IDEAS

    as
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