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Structure paper

TitleStructures of the CRISPR genome integration complex.
Journal, issue, pagesScience, Vol. 357, Issue 6356, Page 1113-1118, Year 2017
Publish dateSep 15, 2017
AuthorsAddison V Wright / Jun-Jie Liu / Gavin J Knott / Kevin W Doxzen / Eva Nogales / Jennifer A Doudna /
PubMed AbstractCRISPR-Cas systems depend on the Cas1-Cas2 integrase to capture and integrate short foreign DNA fragments into the CRISPR locus, enabling adaptation to new viruses. We present crystal structures of ...CRISPR-Cas systems depend on the Cas1-Cas2 integrase to capture and integrate short foreign DNA fragments into the CRISPR locus, enabling adaptation to new viruses. We present crystal structures of Cas1-Cas2 bound to both donor and target DNA in intermediate and product integration complexes, as well as a cryo-electron microscopy structure of the full CRISPR locus integration complex, including the accessory protein IHF (integration host factor). The structures show unexpectedly that indirect sequence recognition dictates integration site selection by favoring deformation of the repeat and the flanking sequences. IHF binding bends the DNA sharply, bringing an upstream recognition motif into contact with Cas1 to increase both the specificity and efficiency of integration. These results explain how the Cas1-Cas2 CRISPR integrase recognizes a sequence-dependent DNA structure to ensure site-selective CRISPR array expansion during the initial step of bacterial adaptive immunity.
External linksScience / PubMed:28729350 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.9 - 3.89 Å
Structure data

8827

5wfe

EMDB-8827, PDB-5wfe:
Cas1-Cas2-IHF-DNA holo-complex
Method: EM (single particle) / Resolution: 3.64 Å

PDB-5vvj:
Cas1-Cas2 bound to half-site intermediate
Method: X-RAY DIFFRACTION / Resolution: 3.89 Å

PDB-5vvk:
Cas1-Cas2 bound to full-site mimic
Method: X-RAY DIFFRACTION / Resolution: 2.9 Å

PDB-5vvl:
Cas1-Cas2 bound to full-site mimic with Ni
Method: X-RAY DIFFRACTION / Resolution: 3.31 Å

Chemicals

ChemComp-NI:
NICKEL (II) ION

Source
  • Escherichia coli (E. coli)
  • escherichia coli k-12 (bacteria)
  • escherichia coli s88 (bacteria)
  • synthetic construct (others)
  • escherichia coli (strain k12) (bacteria)
KeywordsHYDROLASE/DNA / Complex / DNA / HYDROLASE-DNA complex / DNA BINDING PROTEIN/DNA / CRISPR integration complex / Cas1-Cas2 / IHF / DNA BINDING PROTEIN / DNA BINDING PROTEIN-DNA complex

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