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 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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 3765-3783 |
| Number of pages | 19 |
| Journal | Nucleic acids research |
| Volume | 47 |
| Issue number | 7 |
| DOIs | |
| State | Published - Jan 1 2019 |
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ASJC Scopus subject areas
- Genetics
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Structural organization of a Type III-A CRISPR effector subcomplex determined by X-ray crystallography and cryo-EM. / Dorsey, Bryan W.; Huang, Lei; Mondragon, Alfonso.
In: Nucleic acids research, Vol. 47, No. 7, 01.01.2019, p. 3765-3783.Research output: Contribution to journal › Article
TY - JOUR
T1 - Structural organization of a Type III-A CRISPR effector subcomplex determined by X-ray crystallography and cryo-EM
AU - Dorsey, Bryan W.
AU - Huang, Lei
AU - Mondragon, Alfonso
PY - 2019/1/1
Y1 - 2019/1/1
N2 - 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.
AB - 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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UR - http://www.scopus.com/inward/citedby.url?scp=85064982525&partnerID=8YFLogxK
U2 - 10.1093/nar/gkz079
DO - 10.1093/nar/gkz079
M3 - Article
C2 - 30759237
AN - SCOPUS:85064982525
VL - 47
SP - 3765
EP - 3783
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 7
ER -