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

Ning,Cui1; Jun-Tao,Zhang1; Zhuolin,Li1; Xiao-Yu,Liu1; Chongyuan,Wang2,3; Hongda,Huang4; Ning,Jia1,5,6

2022-12-07

10.1038/s41467-022-35275-5

NATURE COMMUNICATIONS   Impact Factor & Quartile

2041-1723

["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.","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 gRAMP(crRNA) and gRAMP(crRNA)-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."]

SCI

English

First ; 共同第一 ; Corresponding

National Natural Science Foundation of China[32270050] ; Shenzhen and Guangdong Natural Science Foundation["202205303000517","2314050005743"] ; Shenzhen Government "Peacock Plan"["Y01226136","Y01416126"] ; Guangdong Provincial Science and Technology Innovation Council[2017B030301018] ; Shenzhen Science and Technology Program[KQTD20190929173906742] ; Key Laboratory of Molecular Design for Plant Cell Factory of Guangdong Higher Education Institutes[2019KSYS006]

Science & Technology - Other Topics

Multidisciplinary Sciences

WOS:000939399900001

NATURE PORTFOLIO

Web of Science

1.Department of Biochemistry, School of Medicine, Southern University of Science and Technology, Shenzhen, 518055, China
2.Faculty of Pharmaceutical Sciences, Shenzhen Institutes of Advanced Technology, Chinese Academy of Science, Shenzhen, 518055, China
3.Center for Human Tissues and Organs Degeneration, Shenzhen Institutes of Advanced Technology, Chinese Academy of Science, Shenzhen, 518055, China
4.Key Laboratory of Molecular Design for Plant Cell Factory of Guangdong Higher Education Institutes, Department of Biology, School of Life Sciences, Southern University of Science and Technology, Shenzhen, 518055, China
5.Shenzhen Key Laboratory of Cell Microenvironment, Guangdong Provincial Key Laboratory of Cell Microenvironment and Disease Research, Southern University of Science and Technology, Shenzhen, 518055, China
6.Key University Laboratory of Metabolism and Health of Guangdong, Southern University of Science and Technology, Shenzhen, 518055, China

Ning,Cui,Jun-Tao,Zhang,Zhuolin,Li,et al. Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase[J]. NATURE COMMUNICATIONS,2022,13(1).

Ning,Cui.,Jun-Tao,Zhang.,Zhuolin,Li.,Xiao-Yu,Liu.,Chongyuan,Wang.,...&Ning,Jia.(2022).Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase.NATURE COMMUNICATIONS,13(1).

Ning,Cui,et al."Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase".NATURE COMMUNICATIONS 13.1(2022).