Cargando…
Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase
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, b...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9729208/ https://www.ncbi.nlm.nih.gov/pubmed/36477448 http://dx.doi.org/10.1038/s41467-022-35275-5 |
_version_ | 1784845436536750080 |
---|---|
author | Cui, Ning Zhang, Jun-Tao Li, Zhuolin Liu, Xiao-Yu Wang, Chongyuan Huang, Hongda Jia, Ning |
author_facet | Cui, Ning Zhang, Jun-Tao Li, Zhuolin Liu, Xiao-Yu Wang, Chongyuan Huang, Hongda Jia, Ning |
author_sort | Cui, Ning |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9729208 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-97292082022-12-09 Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase Cui, Ning Zhang, Jun-Tao Li, Zhuolin Liu, Xiao-Yu Wang, Chongyuan Huang, Hongda Jia, Ning Nat Commun Article 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. Nature Publishing Group UK 2022-12-07 /pmc/articles/PMC9729208/ /pubmed/36477448 http://dx.doi.org/10.1038/s41467-022-35275-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Cui, Ning Zhang, Jun-Tao Li, Zhuolin Liu, Xiao-Yu Wang, Chongyuan Huang, Hongda Jia, Ning Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase |
title | Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase |
title_full | Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase |
title_fullStr | Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase |
title_full_unstemmed | Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase |
title_short | Structural basis for the non-self RNA-activated protease activity of the type III-E CRISPR nuclease-protease Craspase |
title_sort | structural basis for the non-self rna-activated protease activity of the type iii-e crispr nuclease-protease craspase |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9729208/ https://www.ncbi.nlm.nih.gov/pubmed/36477448 http://dx.doi.org/10.1038/s41467-022-35275-5 |
work_keys_str_mv | AT cuining structuralbasisforthenonselfrnaactivatedproteaseactivityofthetypeiiiecrisprnucleaseproteasecraspase AT zhangjuntao structuralbasisforthenonselfrnaactivatedproteaseactivityofthetypeiiiecrisprnucleaseproteasecraspase AT lizhuolin structuralbasisforthenonselfrnaactivatedproteaseactivityofthetypeiiiecrisprnucleaseproteasecraspase AT liuxiaoyu structuralbasisforthenonselfrnaactivatedproteaseactivityofthetypeiiiecrisprnucleaseproteasecraspase AT wangchongyuan structuralbasisforthenonselfrnaactivatedproteaseactivityofthetypeiiiecrisprnucleaseproteasecraspase AT huanghongda structuralbasisforthenonselfrnaactivatedproteaseactivityofthetypeiiiecrisprnucleaseproteasecraspase AT jianing structuralbasisforthenonselfrnaactivatedproteaseactivityofthetypeiiiecrisprnucleaseproteasecraspase |