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TitleMechanisms for HNH-mediated target DNA cleavage in type I CRISPR-Cas systems.
Journal, issue, pagesMol Cell, Vol. 84, Issue 16, Page 3141-33153.e5, Year 2024
Publish dateAug 22, 2024
AuthorsChendi Zhang / Fugen Chen / Feng Wang / Haijiang Xu / Jialin Xue / Zhuang Li /
PubMed AbstractThe metagenome-derived type I-E and type I-F variant CRISPR-associated complex for antiviral defense (Cascade) complexes, fused with HNH domains, precisely cleave target DNA, representing recently ...The metagenome-derived type I-E and type I-F variant CRISPR-associated complex for antiviral defense (Cascade) complexes, fused with HNH domains, precisely cleave target DNA, representing recently identified genome editing tools. However, the underlying working mechanisms remain unknown. Here, structures of type I-F and I-E Cascade complexes at different states are reported. In type I-F Cascade, Cas8f loosely attaches to Cascade head and is adjacent to the 5' end of the target single-stranded DNA (ssDNA). Formation of the full R-loop drives the Cascade head to move outward, allowing Cas8f to detach and rotate ∼150° to accommodate target ssDNA for cleavage. In type I-E Cascade, Cas5e domain is adjacent to the 5' end of the target ssDNA. Full crRNA-target pairing drives the lift of the Cascade head, widening the substrate channel for target ssDNA entrance. Altogether, these analyses into both complexes revealed that crRNA-guided positioning of target DNA and target DNA-induced HNH unlocking are two key factors for their site-specific cleavage of target DNA.
External linksMol Cell / PubMed:39047725
MethodsEM (single particle)
Resolution3.06 - 3.44 Å
Structure data

39110

8yb6

EMDB-39110, PDB-8yb6:
Type I-EHNH Cascade complex
Method: EM (single particle) / Resolution: 3.06 Å

39167

8ydb

EMDB-39167, PDB-8ydb:
Type I-FHNH Cascade-dsDNA intermediate complex
Method: EM (single particle) / Resolution: 3.4 Å

39200

8yeo

EMDB-39200, PDB-8yeo:
Type I-FHNH Cascade-dsDNA R-loop complex
Method: EM (single particle) / Resolution: 3.44 Å

39285

8yh9

EMDB-39285, PDB-8yh9:
Type I-FHNH Cascade complex
Method: EM (single particle) / Resolution: 3.35 Å

39286

8yha

EMDB-39286, PDB-8yha:
Type I-EHNH Cascade-ssDNA complex
Method: EM (single particle) / Resolution: 3.4 Å

Chemicals

ChemComp-ZN:
Unknown entry

Source
  • candidatus cloacimonetes bacterium adurb.bin088 (bacteria)
  • selenomonas sp. (bacteria)
  • candidatus cloacimonadota bacterium (bacteria)
KeywordsRNA BINDING PROTEIN/RNA / RNA pro / RNA BINDING PROTEIN-RNA complex / RNA BINDING PROTEIN/DNA/RNA / protein / RNA / RNA BINDING PROTEIN-DNA-RNA complex

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