热球菌属 Thermus thermophilus 的 RNA 靶向型 III-B CRISPR-Cas 复合物的结构与活性。
Structure and activity of the RNA-targeting Type III-B CRISPR-Cas complex of Thermus thermophilus.
机构信息
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, 6703 HB Wageningen, The Netherlands.
RIKEN SPring-8 Center, 679-5148 Hyogo, Japan.
出版信息
Mol Cell. 2013 Oct 10;52(1):135-145. doi: 10.1016/j.molcel.2013.09.013.
Abstract
The CRISPR-Cas system is a prokaryotic host defense system against genetic elements. The Type III-B CRISPR-Cas system of the bacterium Thermus thermophilus, the TtCmr complex, is composed of six different protein subunits (Cmr1-6) and one crRNA with a stoichiometry of Cmr112131445361:crRNA1. The TtCmr complex copurifies with crRNA species of 40 and 46 nt, originating from a distinct subset of CRISPR loci and spacers. The TtCmr complex cleaves the target RNA at multiple sites with 6 nt intervals via a 5' ruler mechanism. Electron microscopy revealed that the structure of TtCmr resembles a "sea worm" and is composed of a Cmr2-3 heterodimer "tail," a helical backbone of Cmr4 subunits capped by Cmr5 subunits, and a curled "head" containing Cmr1 and Cmr6. Despite having a backbone of only four Cmr4 subunits and being both longer and narrower, the overall architecture of TtCmr resembles that of Type I Cascade complexes.
摘要
CRISPR-Cas 系统是原核生物抵御遗传元件的宿主防御系统。嗜热栖热菌的 III-B 型 CRISPR-Cas 系统，即 TtCmr 复合物，由六个不同的蛋白亚基（Cmr1-6）和一个 crRNA 组成，其摩尔比为 Cmr112131445361:crRNA1。TtCmr 复合物与 40 和 46 nt 的 crRNA 种类共同纯化，这些 crRNA 来源于不同的 CRISPR 基因座和间隔区。TtCmr 复合物通过 5' 规则机制在多个位置以 6 个核苷酸的间隔切割靶 RNA。电子显微镜显示，TtCmr 的结构类似于“海虫”，由 Cmr2-3 异二聚体“尾巴”、Cmr4 亚基的螺旋骨架以及包含 Cmr1 和 Cmr6 的卷曲“头部”组成。尽管只有四个 Cmr4 亚基组成的骨架，且更长更窄，TtCmr 的整体结构类似于 I 型 Casade 复合物。
相似文献
1
Structure and activity of the RNA-targeting Type III-B CRISPR-Cas complex of Thermus thermophilus.热球菌属 Thermus thermophilus 的 RNA 靶向型 III-B CRISPR-Cas 复合物的结构与活性。
Mol Cell. 2013 Oct 10;52(1):135-145. doi: 10.1016/j.molcel.2013.09.013.
2
RNA targeting by the type III-A CRISPR-Cas Csm complex of Thermus thermophilus.嗜热栖热菌III-A型CRISPR-Cas Csm复合物对RNA的靶向作用
Mol Cell. 2014 Nov 20;56(4):518-30. doi: 10.1016/j.molcel.2014.10.005. Epub 2014 Nov 6.
3
Structure of an RNA silencing complex of the CRISPR-Cas immune system.CRISPR-Cas 免疫系统的 RNA 沉默复合物的结构。
Mol Cell. 2013 Oct 10;52(1):146-52. doi: 10.1016/j.molcel.2013.09.008.
4
Target RNA capture and cleavage by the Cmr type III-B CRISPR-Cas effector complex.Cmr III-B型CRISPR-Cas效应复合物对靶RNA的捕获与切割
Genes Dev. 2014 Nov 1;28(21):2432-43. doi: 10.1101/gad.250712.114.
5
CRISPR RNA and anti-CRISPR protein binding to the Csy1-Csy2 heterodimer in the type I-F CRISPR-Cas system.I-F 型 CRISPR-Cas 系统中 CRISPR RNA 与抗 CRISPR 蛋白结合到 Csy1-Csy2 异二聚体上。
J Biol Chem. 2018 Feb 23;293(8):2744-2754. doi: 10.1074/jbc.RA117.001611. Epub 2018 Jan 18.
6
RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes.功能正交的VI-A型CRISPR-Cas酶对RNA的靶向作用
Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
7
Type III CRISPR complexes from Thermus thermophilus.嗜热栖热菌的III型CRISPR复合体。
Acta Biochim Pol. 2016;63(2):377-86. doi: 10.18388/abp.2016_1261. Epub 2016 Jun 14.
8
Cmr4 is the slicer in the RNA-targeting Cmr CRISPR complex.Cmr4是RNA靶向Cmr CRISPR复合物中的切割酶。
Nucleic Acids Res. 2015 Jan;43(2):1257-67. doi: 10.1093/nar/gku1355. Epub 2014 Dec 24.
9
Type III-A CRISPR-Cas Csm Complexes: Assembly, Periodic RNA Cleavage, DNase Activity Regulation, and Autoimmunity.III-A 型 CRISPR-Cas Csm 复合物：组装、周期性 RNA 切割、DNase 活性调节和自身免疫。
Mol Cell. 2019 Jan 17;73(2):264-277.e5. doi: 10.1016/j.molcel.2018.11.007. Epub 2018 Nov 29.
10
Crystal structure of the CRISPR-Cas RNA silencing Cmr complex bound to a target analog.CRISPR-Cas RNA 沉默 Cmr 复合物与靶类似物结合的晶体结构。
Mol Cell. 2015 May 7;58(3):418-30. doi: 10.1016/j.molcel.2015.03.018. Epub 2015 Apr 23.
引用本文的文献
1
CRISPR Cas6 Heterologous Expression and Purification.CRISPR Cas6的异源表达与纯化。
Bio Protoc. 2025 Jul 20;15(14):e5382. doi: 10.21769/BioProtoc.5382.
2
A type III-associated Cas6 functions as a negative regulator of type I-B CRISPR-Cas system in Thermus thermophilus.III型相关的Cas6在嗜热栖热菌中作为I-B型CRISPR-Cas系统的负调控因子发挥作用。
Commun Biol. 2025 May 23;8(1):793. doi: 10.1038/s42003-025-08223-4.
3
The influence of the copy number of invader on the fate of bacterial host cells in the antiviral defense by CRISPR-Cas10 DNases.在CRISPR-Cas10脱氧核糖核酸酶介导的抗病毒防御中，入侵核酸拷贝数对细菌宿主细胞命运的影响。
Eng Microbiol. 2023 Jun 24;3(4):100102. doi: 10.1016/j.engmic.2023.100102. eCollection 2023 Dec.
4
AcrIIIA1 is a protein-RNA anti-CRISPR complex that targets core Cas and accessory nucleases.AcrIIIA1是一种蛋白质-RNA抗CRISPR复合物，其靶向核心Cas和辅助核酸酶。
Nucleic Acids Res. 2024 Dec 11;52(22):13490-13514. doi: 10.1093/nar/gkae1006.
5
Selective degradation of phage RNAs by the Csm6 ribonuclease provides robust type III CRISPR immunity in Streptococcus thermophilus.Csm6 核糖核酸酶对噬菌体 RNA 的选择性降解为嗜热链球菌提供了强大的 III 型 CRISPR 免疫。
Nucleic Acids Res. 2024 Nov 11;52(20):12549-12564. doi: 10.1093/nar/gkae856.
6
Structural basis for the type I-F Cas8-HNH system.I-F Cas8-HNH 系统的结构基础。
EMBO J. 2024 Oct;43(20):4656-4667. doi: 10.1038/s44318-024-00229-8. Epub 2024 Sep 9.
7
Nanopores Reveal the Stoichiometry of Single Oligoadenylates Produced by Type III CRISPR-Cas.纳米孔揭示了 III 型 CRISPR-Cas 产生的单聚腺苷酸的化学计量。
ACS Nano. 2024 Jul 2;18(26):16505-16515. doi: 10.1021/acsnano.3c11769. Epub 2024 Jun 14.
8
High-efficiency genome editing of an extreme thermophile using endogenous type I and type III CRISPR-Cas systems.利用内源性I型和III型CRISPR-Cas系统对嗜热放线菌进行高效基因组编辑。
mLife. 2022 Dec 7;1(4):412-427. doi: 10.1002/mlf2.12045. eCollection 2022 Dec.
9
RNA targeting and cleavage by the type III-Dv CRISPR effector complex.III-Dv 型 CRISPR 效应物复合物对 RNA 的靶向和切割。
Nat Commun. 2024 Apr 18;15(1):3324. doi: 10.1038/s41467-024-47506-y.
10
Retention of the RNA ends provides the molecular memory for maintaining the activation of the Csm complex.RNA末端的保留为维持Csm复合物的激活提供了分子记忆。
Nucleic Acids Res. 2024 Apr 24;52(7):3896-3910. doi: 10.1093/nar/gkae080.
本文引用的文献
1
A ruler protein in a complex for antiviral defense determines the length of small interfering CRISPR RNAs.一种抗病毒防御复合物中的调控蛋白决定了小干扰 CRISPR RNA 的长度。
J Biol Chem. 2013 Sep 27;288(39):27888-97. doi: 10.1074/jbc.M113.499244. Epub 2013 Aug 9.
2
CRISPR interference: a structural perspective.CRISPR 干扰：结构视角。
Biochem J. 2013 Jul 15;453(2):155-66. doi: 10.1042/BJ20130316.
3
Crystal structure of the Cmr2-Cmr3 subcomplex in the CRISPR-Cas RNA silencing effector complex.CRISPR-Cas RNA 沉默效应物复合物中 Cmr2-Cmr3 亚基复合物的晶体结构。
J Mol Biol. 2013 Oct 23;425(20):3811-23. doi: 10.1016/j.jmb.2013.03.042. Epub 2013 Apr 10.
4
CRISPR-Cas systems in the cyanobacterium Synechocystis sp. PCC6803 exhibit distinct processing pathways involving at least two Cas6 and a Cmr2 protein.集胞藻 PCC6803 中的 CRISPR-Cas 系统表现出不同的加工途径，涉及至少两种 Cas6 和一种 Cmr2 蛋白。
PLoS One. 2013;8(2):e56470. doi: 10.1371/journal.pone.0056470. Epub 2013 Feb 18.
5
Structure of the Cmr2-Cmr3 subcomplex of the Cmr RNA silencing complex.Cmr RNA 沉默复合物的 Cmr2-Cmr3 亚复合物结构。
Structure. 2013 Mar 5;21(3):376-84. doi: 10.1016/j.str.2013.01.002. Epub 2013 Feb 7.
6
A novel interference mechanism by a type IIIB CRISPR-Cmr module in Sulfolobus.一种新型的 IIIB 型 CRISPR-Cmr 模块在 Sulfolobus 中的干扰机制。
Mol Microbiol. 2013 Mar;87(5):1088-99. doi: 10.1111/mmi.12152. Epub 2013 Feb 3.
7
Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria.Cas9-crRNA 核糖核蛋白复合物介导细菌适应性免疫中的特异性 DNA 切割。
Proc Natl Acad Sci U S A. 2012 Sep 25;109(39):E2579-86. doi: 10.1073/pnas.1208507109. Epub 2012 Sep 4.
8
Native tandem and ion mobility mass spectrometry highlight structural and modular similarities in clustered-regularly-interspaced shot-palindromic-repeats (CRISPR)-associated protein complexes from Escherichia coli and Pseudomonas aeruginosa.天然串联和离子淌度质谱突出了大肠杆菌和铜绿假单胞菌中聚类规则间隔短回文重复（CRISPR）相关蛋白复合物的结构和模块相似性。
Mol Cell Proteomics. 2012 Nov;11(11):1430-41. doi: 10.1074/mcp.M112.020263. Epub 2012 Aug 23.
9
Cas5d protein processes pre-crRNA and assembles into a cascade-like interference complex in subtype I-C/Dvulg CRISPR-Cas system.Cas5d 蛋白加工前体 crRNA 并在 I-C/Dvulg CRISPR-Cas 系统的亚类 I 中组装成级联样干扰复合物。
Structure. 2012 Sep 5;20(9):1574-84. doi: 10.1016/j.str.2012.06.016. Epub 2012 Jul 26.
10
A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.可编程的双 RNA 引导的 DNA 内切酶在适应性细菌免疫中的作用。
Science. 2012 Aug 17;337(6096):816-21. doi: 10.1126/science.1225829. Epub 2012 Jun 28.
Zotero 插件