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Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081

The cyclic oligoadenylates (cOAs) act as second messengers of the type III CRISPR immunity system through activating the auxiliary nucleases for indiscriminate RNA degradation. The cOA-degrading nucleases (ring nucleases) provide an ‘off-switch’ regulation of the signaling, thereby preventing cell d...

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Detalles Bibliográficos
Autores principales: Du, Liyang, Zhang, Danping, Luo, Zhipu, Lin, Zhonghui
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10018336/
https://www.ncbi.nlm.nih.gov/pubmed/36807980
http://dx.doi.org/10.1093/nar/gkad101
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author Du, Liyang
Zhang, Danping
Luo, Zhipu
Lin, Zhonghui
author_facet Du, Liyang
Zhang, Danping
Luo, Zhipu
Lin, Zhonghui
author_sort Du, Liyang
collection PubMed
description The cyclic oligoadenylates (cOAs) act as second messengers of the type III CRISPR immunity system through activating the auxiliary nucleases for indiscriminate RNA degradation. The cOA-degrading nucleases (ring nucleases) provide an ‘off-switch’ regulation of the signaling, thereby preventing cell dormancy or cell death. Here, we describe the crystal structures of the founding member of CRISPR-associated ring nuclease 1 (Crn1) Sso2081 from Saccharolobus solfataricus, alone, bound to phosphate ions or cA(4) in both pre-cleavage and cleavage intermediate states. These structures together with biochemical characterizations establish the molecular basis of cA(4) recognition and catalysis by Sso2081. The conformational changes in the C-terminal helical insert upon the binding of phosphate ions or cA(4) reveal a gate-locking mechanism for ligand binding. The critical residues and motifs identified in this study provide a new insight to distinguish between cOA-degrading and -nondegrading CARF domain-containing proteins.
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spelling pubmed-100183362023-03-17 Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081 Du, Liyang Zhang, Danping Luo, Zhipu Lin, Zhonghui Nucleic Acids Res Structural Biology The cyclic oligoadenylates (cOAs) act as second messengers of the type III CRISPR immunity system through activating the auxiliary nucleases for indiscriminate RNA degradation. The cOA-degrading nucleases (ring nucleases) provide an ‘off-switch’ regulation of the signaling, thereby preventing cell dormancy or cell death. Here, we describe the crystal structures of the founding member of CRISPR-associated ring nuclease 1 (Crn1) Sso2081 from Saccharolobus solfataricus, alone, bound to phosphate ions or cA(4) in both pre-cleavage and cleavage intermediate states. These structures together with biochemical characterizations establish the molecular basis of cA(4) recognition and catalysis by Sso2081. The conformational changes in the C-terminal helical insert upon the binding of phosphate ions or cA(4) reveal a gate-locking mechanism for ligand binding. The critical residues and motifs identified in this study provide a new insight to distinguish between cOA-degrading and -nondegrading CARF domain-containing proteins. Oxford University Press 2023-02-20 /pmc/articles/PMC10018336/ /pubmed/36807980 http://dx.doi.org/10.1093/nar/gkad101 Text en © The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Structural Biology
Du, Liyang
Zhang, Danping
Luo, Zhipu
Lin, Zhonghui
Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081
title Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081
title_full Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081
title_fullStr Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081
title_full_unstemmed Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081
title_short Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081
title_sort molecular basis of stepwise cyclic tetra-adenylate cleavage by the type iii crispr ring nuclease crn1/sso2081
topic Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10018336/
https://www.ncbi.nlm.nih.gov/pubmed/36807980
http://dx.doi.org/10.1093/nar/gkad101
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