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TitleStructural organization of a Type III-A CRISPR effector subcomplex determined by X-ray crystallography and cryo-EM.
Journal, issue, pagesNucleic Acids Res, Vol. 47, Issue 7, Page 3765-3783, Year 2019
Publish dateApr 23, 2019
AuthorsBryan W Dorsey / Lei Huang / Alfonso Mondragón /
PubMed AbstractClustered regularly interspaced short palindromic repeats (CRISPR) and their associated Cas proteins provide an immune-like response in many prokaryotes against extraneous nucleic acids. CRISPR-Cas ...Clustered regularly interspaced short palindromic repeats (CRISPR) and their associated Cas proteins provide an immune-like response in many prokaryotes against extraneous nucleic acids. CRISPR-Cas systems are classified into different classes and types. Class 1 CRISPR-Cas systems form multi-protein effector complexes that includes a guide RNA (crRNA) used to identify the target for destruction. Here we present crystal structures of Staphylococcus epidermidis Type III-A CRISPR subunits Csm2 and Csm3 and a 5.2 Å resolution single-particle cryo-electron microscopy (cryo-EM) reconstruction of an in vivo assembled effector subcomplex including the crRNA. The structures help to clarify the quaternary architecture of Type III-A effector complexes, and provide details on crRNA binding, target RNA binding and cleavage, and intermolecular interactions essential for effector complex assembly. The structures allow a better understanding of the organization of Type III-A CRISPR effector complexes as well as highlighting the overall similarities and differences with other Class 1 effector complexes.
External linksNucleic Acids Res / PubMed:30759237 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.4 - 5.2 Å
Structure data

EMDB-0442:
Type III-A CRISPR Effector Subcomplex from Staphylococcus epidermidis RP62a
Method: EM (single particle) / Resolution: 5.2 Å

PDB-6nbt:
CRISPR Complex Subunit Csm3 from Staphylococcus epidermidis RP62a
Method: X-RAY DIFFRACTION / Resolution: 2.4 Å

PDB-6nbu:
CRISPR Complex Subunit Csm2 from Staphylococcus epidermidis RP62a
Method: X-RAY DIFFRACTION / Resolution: 2.75 Å

Chemicals

ChemComp-CA:
Unknown entry

ChemComp-SM:
SAMARIUM (III) ION

ChemComp-HOH:
WATER

Source
  • Staphylococcus epidermidis RP62A (bacteria)
  • staphylococcus epidermidis (strain atcc 35984 / rp62a) (bacteria)
KeywordsRNA BINDING PROTEIN / CRISPR / RNA Binding / RNA Recognition Motif / Samarium (III) chloride / STRUCTURAL PROTEIN

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