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TitleStructure of the DNA-Bound Spacer Capture Complex of a Type II CRISPR-Cas System.
Journal, issue, pagesMol Cell, Vol. 75, Issue 1, Page 90-101.e5, Year 2019
Publish dateJul 11, 2019
AuthorsMartin Wilkinson / Gediminas Drabavicius / Arunas Silanskas / Giedrius Gasiunas / Virginijus Siksnys / Dale B Wigley /
PubMed AbstractCRISPR and associated Cas proteins function as an adaptive immune system in prokaryotes to combat bacteriophage infection. During the immunization step, new spacers are acquired by the CRISPR ...CRISPR and associated Cas proteins function as an adaptive immune system in prokaryotes to combat bacteriophage infection. During the immunization step, new spacers are acquired by the CRISPR machinery, but the molecular mechanism of spacer capture remains enigmatic. We show that the Cas9, Cas1, Cas2, and Csn2 proteins of a Streptococcus thermophilus type II-A CRISPR-Cas system form a complex and provide cryoelectron microscopy (cryo-EM) structures of three different assemblies. The predominant form, with the stoichiometry Cas1-Cas2-Csn2, referred to as monomer, contains ∼30 bp duplex DNA bound along a central channel. A minor species, termed a dimer, comprises two monomers that sandwich a further eight Cas1 and four Cas2 subunits and contains two DNA ∼30-bp duplexes within the channel. A filamentous form also comprises Cas1-Cas2-Csn2 units (typically 2-6) but with a different Cas1-Cas2 interface between them and a continuous DNA duplex running along a central channel.
External linksMol Cell / PubMed:31080012 / PubMed Central
MethodsEM (single particle)
Resolution3.6 - 9.1 Å
Structure data

4668

6qxf

EMDB-4668, PDB-6qxf:
Cas1-Cas2-Csn2-DNA complex from the Type II-A CRISPR-Cas system
Method: EM (single particle) / Resolution: 3.6 Å

4670

6qxt

EMDB-4670, PDB-6qxt:
Cas1-Cas2-Csn2-DNA dimer complex from the Type II-A CRISPR-Cas system
Method: EM (single particle) / Resolution: 8.9 Å

4671

6qy3

EMDB-4671, PDB-6qy3:
Segment of the Cas1-Cas2-Csn2-DNA filament complex from the Type II-A CRISPR-Cas system
Method: EM (single particle) / Resolution: 9.1 Å

Chemicals

ChemComp-CA:
Unknown entry

Source
  • streptococcus thermophilus (bacteria)
  • escherichia coli bl21(de3) (bacteria)
KeywordsDNA BINDING PROTEIN / CRISPR / Cas / DNA binding / Nuclease / Adaptation / Spacer acquisition / Protein complex / Protein-DNA complex / Calcium / Metal-binding / Antiviral

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