Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081.
Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081.
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DOI：
10.1093/nar/gkad101
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发表时间：
2023-03-21
影响因子：
14.9
通讯作者：
中科院分区：
文献类型：
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作者：
环状寡聚腺苷酸（cOA）通过激活辅助核酸酶来无差别地降解RNA，从而充当III型CRISPR免疫系统的第二信使。降解cOA的核酸酶（环核酸酶）提供信号传导的“关闭开关”调节，从而防止细胞休眠或细胞死亡。在这里，我们描述了来自Solfataricus的CRISPR相关环核酸酶1（Crn 1）Sso 2081的创始成员的晶体结构，该成员单独在预切割和切割中间状态下与磷酸根离子或cA 4结合。这些结构与生化表征一起建立了Sso 2081识别和催化cA 4的分子基础。磷酸根离子或cA 4结合后C-末端螺旋插入物的构象变化揭示了配体结合的门锁定机制。在这项研究中确定的关键残基和基序提供了一个新的见解，以区分cOA降解和非降解CARF结构域的蛋白质。
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 cA4 in both pre-cleavage and cleavage intermediate states. These structures together with biochemical characterizations establish the molecular basis of cA4 recognition and catalysis by Sso2081. The conformational changes in the C-terminal helical insert upon the binding of phosphate ions or cA4 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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DOI：
10.1126/science.1165771
发表时间：
2008-12-19
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Marraffini LA;Sontheimer EJ
通讯作者：
Sontheimer EJ
影响因子：
6.8
作者：
Molina, Rafael;Sofos, Nicholas;Montoya, Guillermo
通讯作者：
Montoya, Guillermo
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
14.9
作者：
Molina, Rafael;Garcia-Martin, Ricardo;Lopez-Mendez, Blanca;Jensen, Anne Louise Gron;Ciges-Tomas, J. Rafael;Marchena-Hurtado, Javier;Stella, Stefano;Montoya, Guillermo
通讯作者：
Montoya, Guillermo
影响因子：
64.8
作者：
Athukoralage JS;Rouillon C;Graham S;Grüschow S;White MF
通讯作者：
White MF