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Title | Structural organization of a Type III-A CRISPR effector subcomplex determined by X-ray crystallography and cryo-EM. |
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Journal, issue, pages | Nucleic Acids Res, Vol. 47, Issue 7, Page 3765-3783, Year 2019 |
Publish date | Apr 23, 2019 |
![]() | Bryan W Dorsey / Lei Huang / Alfonso Mondragón / ![]() |
PubMed Abstract | 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 ...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. |
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Methods | EM (single particle) / X-ray diffraction |
Resolution | 2.4 - 5.2 Å |
Structure data | ![]() EMDB-0442: ![]() PDB-6nbt: ![]() PDB-6nbu: |
Chemicals | ![]() ChemComp-CA: ![]() ChemComp-SM: ![]() ChemComp-HOH: |
Source |
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![]() | RNA BINDING PROTEIN / CRISPR / RNA Binding / RNA Recognition Motif / Samarium (III) chloride / STRUCTURAL PROTEIN |