Structure of the OMEGA nickase IsrB in complex with ωRNA and target DNA.
Structure of the OMEGA nickase IsrB in complex with ωRNA and target DNA.
复制标题
DOI：
10.1038/s41586-022-05324-6
复制
发表时间：
2022-10
期刊：
影响因子：
64.8
通讯作者：
Zhang, Feng
中科院分区：
文献类型：
--
作者：
Hirano, Seiichi;Kappel, Kalli;Altae-Tran, Han;Faure, Guilhem;Wilkinson, Max E.;Kannan, Soumya;Demircioglu, F. Esra;Yan, Rui;Shiozaki, Momoko;Yu, Zhiheng;Makarova, Kira S.;Koonin, Eugene, V;Macrae, Rhiannon K.;Zhang, Feng
RNA引导系统，如CRISPR-Cas，将可编程底物识别与酶功能相结合，这种组合已被有利地用于开发强大的分子技术。对这些系统的结构研究阐明了RNA和蛋白质如何共同识别和切割其底物，为进一步的技术开发指导合理的工程设计。最近的工作确定了一类新的RNA引导系统，称为OMEGA，其中包括IscB（Cas9的可能祖先）和切口酶IsrB（缺乏HNH核酸酶结构域的IscB同源物）。IsrB仅由大约350个氨基酸组成，但它的小尺寸被相对较大的RNA向导（大约300-nt ωRNA）所抵消。在这里，我们报告的低温电子显微镜结构的Desulfovirgula thermocuniculi IsrB（DtIsrB）在其同源ωRNA和靶DNA的复合物。我们发现IsrB蛋白的整体结构与Cas9共享一个共同的支架。然而，与Cas9相比，Cas9使用识别（REC）叶来促进靶点选择，IsrB依赖于其ωRNA，其中一部分形成了类似于REC的复杂三元结构。IsrB及其ωRNA的结构分析以及与其他RNA引导系统的比较突出了蛋白质和RNA之间的功能相互作用，促进了我们对这些不同系统的生物学和进化的理解。低温电子显微镜下观察到的D.热兔IsrB蛋白与其同源ωRNA和靶DNA的复合显示RNA-显性IsrB效应物复合物与蛋白质-显性Cas9效应物复合物共享共同的支架。
RNA-guided systems, such as CRISPR–Cas, combine programmable substrate recognition with enzymatic function, a combination that has been used advantageously to develop powerful molecular technologies. Structural studies of these systems have illuminated how the RNA and protein jointly recognize and cleave their substrates, guiding rational engineering for further technology development. Recent work identified a new class of RNA-guided systems, termed OMEGA, which include IscB, the likely ancestor of Cas9, and the nickase IsrB, a homologue of IscB lacking the HNH nuclease domain. IsrB consists of only around 350 amino acids, but its small size is counterbalanced by a relatively large RNA guide (roughly 300-nt ωRNA). Here, we report the cryogenic-electron microscopy structure of Desulfovirgula thermocuniculi IsrB (DtIsrB) in complex with its cognate ωRNA and a target DNA. We find the overall structure of the IsrB protein shares a common scaffold with Cas9. In contrast to Cas9, however, which uses a recognition (REC) lobe to facilitate target selection, IsrB relies on its ωRNA, part of which forms an intricate ternary structure positioned analogously to REC. Structural analyses of IsrB and its ωRNA as well as comparisons to other RNA-guided systems highlight the functional interplay between protein and RNA, advancing our understanding of the biology and evolution of these diverse systems. The cryogenic-electron microscopy structure of the D. thermocuniculi IsrB protein in complex with its cognate ωRNA and a target DNA shows that the RNA-dominant IsrB effector complex shares a common scaffold with the protein-dominant Cas9 effector complex.
登录
查看更多内容
地狱1.1：RNA同源性搜索速度更快100倍。
DOI：
10.1093/bioinformatics/btt509
发表时间：
2013-11-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Nawrocki EP;Eddy SR
通讯作者：
Eddy SR
影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
DOI：
10.1042/bcj20210708
发表时间：
2021-12-22
期刊：
The Biochemical journal
影响因子：
--
作者：
Kimanius D;Dong L;Sharov G;Nakane T;Scheres SHW
通讯作者：
Scheres SHW
影响因子：
64.8
作者：
Reiter NJ;Osterman A;Torres-Larios A;Swinger KK;Pan T;Mondragón A
通讯作者：
Mondragón A
影响因子：
3.2
作者：
Kapitonov VV;Makarova KS;Koonin EV
通讯作者：
Koonin EV