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TitleStructure basis for RNA-guided DNA degradation by Cascade and Cas3.
Journal, issue, pagesScience, Vol. 361, Issue 6397, Year 2018
Publish dateJul 6, 2018
AuthorsYibei Xiao / Min Luo / Adam E Dolan / Maofu Liao / Ailong Ke /
PubMed AbstractType I CRISPR-Cas system features a sequential target-searching and degradation process on double-stranded DNA by the RNA-guided Cascade (CRISPR associated complex for antiviral defense) complex and ...Type I CRISPR-Cas system features a sequential target-searching and degradation process on double-stranded DNA by the RNA-guided Cascade (CRISPR associated complex for antiviral defense) complex and the nuclease-helicase fusion enzyme Cas3, respectively. Here, we present a 3.7-angstrom-resolution cryo-electron microscopy (cryo-EM) structure of the Type I-E Cascade/R-loop/Cas3 complex, poised to initiate DNA degradation. Cas3 distinguishes Cascade conformations and only captures the R-loop-forming Cascade, to avoid cleaving partially complementary targets. Its nuclease domain recruits the nontarget strand (NTS) DNA at a bulged region for the nicking of single-stranded DNA. An additional 4.7-angstrom-resolution cryo-EM structure captures the postnicking state, in which the severed NTS retracts to the helicase entrance, to be threaded for adenosine 5'-triphosphate-dependent processive degradation. These snapshots form the basis for understanding RNA-guided DNA degradation in Type I-E CRISPR-Cas systems.
External linksScience / PubMed:29880725 / PubMed Central
MethodsEM (single particle)
Resolution3.66 Å
Structure data

7347

6c66

EMDB-7347, PDB-6c66:
CRISPR RNA-guided surveillance complex, pre-nicking
Method: EM (single particle) / Resolution: 3.66 Å

Chemicals

ChemComp-FE:
Unknown entry

Source
  • thermobifida fusca (strain yx) (bacteria)
  • thermobifida fusca (bacteria)
KeywordsDNA BINDING PROTEIN/DNA/RNA / CRISPR-Cas / Cascade / Cas3 / DNA BINDING PROTEIN / DNA BINDING PROTEIN-DNA-RNA complex

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