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TitleType III-A CRISPR-Cas Csm Complexes: Assembly, Periodic RNA Cleavage, DNase Activity Regulation, and Autoimmunity.
Journal, issue, pagesMol Cell, Vol. 73, Issue 2, Page 264-277.e5, Year 2019
Publish dateJan 17, 2019
AuthorsNing Jia / Charlie Y Mo / Chongyuan Wang / Edward T Eng / Luciano A Marraffini / Dinshaw J Patel /
PubMed AbstractType 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.
External linksMol Cell / PubMed:30503773 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.91 - 3.6 Å
Structure data

9253

6mur

EMDB-9253, PDB-6mur:
Cryo-EM structure of Csm-crRNA-target RNA ternary complex in type III-A CRISPR-Cas system
Method: EM (single particle) / Resolution: 3.1 Å

9254

6mus

EMDB-9254, PDB-6mus:
Cryo-EM structure of larger Csm-crRNA-target RNA ternary complex in type III-A CRISPR-Cas system
Method: EM (single particle) / Resolution: 3.6 Å

9255

6mut

EMDB-9255, PDB-6mut:
Cryo-EM structure of ternary Csm-crRNA-target RNA with anti-tag sequence complex in type III-A CRISPR-Cas system
Method: EM (single particle) / Resolution: 3.1 Å

9256

6muu

EMDB-9256, PDB-6muu:
Cryo-EM structure of Csm-crRNA binary complex in type III-A CRISPR-Cas system
Method: EM (single particle) / Resolution: 3.0 Å

PDB-6mua:
Crystal structure of Csm1-Csm4 subcomplex in the type III-A CRISPR-Csm interference complex
Method: X-RAY DIFFRACTION / Resolution: 2.91 Å

Chemicals

ChemComp-ZN:
Unknown entry

Source
  • thermococcus onnurineus (archaea)
  • thermococcus onnurineus (strain na1) (archaea)
KeywordsIMMUNE 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

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