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TitleStructures of an active type III-A CRISPR effector complex.
Journal, issue, pagesStructure, Vol. 30, Issue 8, Page 1109-11128.e6, Year 2022
Publish dateAug 4, 2022
AuthorsEric M Smith / Sé Ferrell / Valerie L Tokars / Alfonso Mondragón /
PubMed AbstractClustered regularly interspaced short palindromic repeats (CRISPR) and their CRISPR-associated proteins (Cas) provide many prokaryotes with an adaptive immune system against invading genetic material. ...Clustered regularly interspaced short palindromic repeats (CRISPR) and their CRISPR-associated proteins (Cas) provide many prokaryotes with an adaptive immune system against invading genetic material. Type III CRISPR systems are unique in that they can degrade both RNA and DNA. In response to invading nucleic acids, they produce cyclic oligoadenylates that act as secondary messengers, activating cellular nucleases that aid in the immune response. Here, we present seven single-particle cryo-EM structures of the type III-A Staphylococcus epidermidis CRISPR effector complex. The structures reveal the intact S. epidermidis effector complex in an apo, ATP-bound, cognate target RNA-bound, and non-cognate target RNA-bound states and illustrate how the effector complex binds and presents crRNA. The complexes bound to target RNA capture the type III-A effector complex in a post-RNA cleavage state. The ATP-bound structures give details about how ATP binds to Cas10 to facilitate cyclic oligoadenylate production.
External linksStructure / PubMed:35714601 / PubMed Central
MethodsEM (single particle)
Resolution3.49 - 4.97 Å
Structure data

EMDB-26920, PDB-7uzw:
Staphylococcus epidermidis RP62a CRISPR effector subcomplex
Method: EM (single particle) / Resolution: 3.55 Å

EMDB-26921, PDB-7uzx:
Staphylococcus epidermidis RP62a CRISPR effector subcomplex with non-self target RNA bound
Method: EM (single particle) / Resolution: 3.49 Å

EMDB-26922, PDB-7uzy:
Staphylococcus epidermidis RP62A CRISPR effector complex with non-self target RNA 2
Method: EM (single particle) / Resolution: 4.05 Å

EMDB-26923, PDB-7uzz:
Staphylococcus epidermidis RP62a CRISPR tall effector complex
Method: EM (single particle) / Resolution: 4.45 Å

EMDB-26924, PDB-7v00:
Staphylococcus epidermidis RP62a CRISPR tall effector complex with bound ATP
Method: EM (single particle) / Resolution: 3.87 Å

EMDB-26925, PDB-7v01:
Staphylococcus epidermidis RP62a CRISPR short effector complex with self RNA target and ATP
Method: EM (single particle) / Resolution: 3.67 Å

EMDB-26926: Staphylococcus epidermidis RP62A CRISPR effector complex with non-self target RNA 1
Method: EM (single particle) / Resolution: 4.06 Å

EMDB-26927, PDB-7v02:
Staphylococcus epidermidis RP62A CRISPR short effector complex
Method: EM (single particle) / Resolution: 4.97 Å

Chemicals

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM / Adenosine triphosphate

Source
  • staphylococcus epidermidis rp62a (bacteria)
  • staphylococcus epidermidis (bacteria)
KeywordsRNA BINDING PROTEIN/RNA / Type IIIA CRISPR / effector complex / RNA BINDING PROTEIN / RNA BINDING PROTEIN-RNA complex / HYDROLASE/RNA / HYDROLASE-RNA complex

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