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- Title
Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase.
- Authors
Cui, Ning; Zhang, Jun-Tao; Li, Zhuolin; Liu, Xiao-Yu; Wang, Chongyuan; Huang, Hongda; Jia, Ning
- 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. The authors report several high-resolution functional snapshots of type III-E nuclease-protease Craspase complexes, revealing the mechanisms underlying target RNA cleavage and non-self RNA activated protease activities; and highlighting the potentials for the development of RNA-guided nuclease-protease Craspase-based tools for biotechnological applications.
- Subjects
CRISPRS; RNA; CASPASES; NUCLEOTIDES; ZINC-finger proteins; PROTEINS; THROMBIN receptors; PROTEOLYTIC enzymes
- Publication
Nature Communications, 2022, Vol 13, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-022-35275-5