+検索条件
-Structure paper
タイトル | Type III-A CRISPR-Cas Csm Complexes: Assembly, Periodic RNA Cleavage, DNase Activity Regulation, and Autoimmunity. |
---|---|
ジャーナル・号・ページ | Mol Cell, Vol. 73, Issue 2, Page 264-277.e5, Year 2019 |
掲載日 | 2019年1月17日 |
著者 | Ning Jia / Charlie Y Mo / Chongyuan Wang / Edward T Eng / Luciano A Marraffini / Dinshaw J Patel / |
PubMed 要旨 | Type ΙΙΙ CRISPR-Cas systems provide robust immunity against foreign RNA and DNA by sequence-specific RNase and target RNA-activated sequence-nonspecific DNase and RNase activities. We report on ...Type ΙΙΙ CRISPR-Cas systems provide robust immunity against foreign RNA and DNA by sequence-specific RNase and target RNA-activated sequence-nonspecific DNase and RNase activities. We report on cryo-EM structures of Thermococcus onnurineus Csm binary, Csm-target RNA and Csm-target RNA ternary complexes in the 3.1 Å range. The topological features of the crRNA 5'-repeat tag explains the 5'-ruler mechanism for defining target cleavage sites, with accessibility of positions -2 to -5 within the 5'-repeat serving as sensors for avoidance of autoimmunity. The Csm3 thumb elements introduce periodic kinks in the crRNA-target RNA duplex, facilitating cleavage of the target RNA with 6-nt periodicity. Key Glu residues within a Csm1 loop segment of Csm adopt a proposed autoinhibitory conformation suggestive of DNase activity regulation. These structural findings, complemented by mutational studies of key intermolecular contacts, provide insights into Csm complex assembly, mechanisms underlying RNA targeting and site-specific periodic cleavage, regulation of DNase cleavage activity, and autoimmunity suppression. |
リンク | Mol Cell / PubMed:30503773 / PubMed Central |
手法 | EM (単粒子) / X線回折 |
解像度 | 2.91 - 3.6 Å |
構造データ | EMDB-9253, PDB-6mur: EMDB-9254, PDB-6mus: EMDB-9255, PDB-6mut: EMDB-9256, PDB-6muu: PDB-6mua: |
化合物 | ChemComp-ZN: |
由来 |
|
キーワード | IMMUNE SYSTEM / Type III-A CRISPR-Cas system / Csm1-Csm4 subcomplex / RNA BINDING PROTEIN/RNA / cryo-EM structure / Csm-crRNA-target RNA ternary complex / Type III CRISPR-Cas systerm / RNA BINDING PROTEIN-RNA complex / larger Csm-crRNA-target RNA ternary complex / ternary Csm-crRNA-target RNA with anti-tag sequence complex / Type III CRISPR-Cas system / Csm-crRNA binary complex |