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PDBsum entry 5f9r

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protein dna_rna ligands links
Hydrolase/DNA/RNA PDB id
5f9r
Jmol PyMol
Contents
Protein chain
1362 a.a.
DNA/RNA
Ligands
SO4 ×3
PDB id:
5f9r
Name: Hydrolase/DNA/RNA
Title: Crystal structure of catalytically-active streptococcus pyog crispr-cas9 in complex with single-guided RNA and double-st primed for target DNA cleavage
Structure: Crispr-associated endonuclease cas9/csn1. Chain: b. Synonym: spycas9. Engineered: yes. RNA (116-mer). Chain: a. Engineered: yes. Other_details: gtp. DNA (30-mer).
Source: Streptococcus pyogenes serotype m1. Organism_taxid: 301447. Gene: cas9, csn1, spy_1046. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes. Streptococcus pyogenes mgas8232. Organism_taxid: 186103. Other_details: RNA was prepared by in vitro transcription w
Resolution:
3.40Å     R-factor:   0.240     R-free:   0.299
Authors: F.Jiang,J.A.Doudna
Key ref: F.Jiang et al. (2016). Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage. Science, 351, 867-871. PubMed id: 26841432 DOI: 10.1126/science.aad8282
Date:
10-Dec-15     Release date:   27-Jan-16    
PROCHECK
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 Headers
 References

Protein chain
Pfam   ArchSchema ?
Q99ZW2  (Q99ZW2_STRP1) -  CRISPR-associated endonuclease Cas9/Csn1
Seq:
Struc:
 
Seq:
Struc:
 
Seq:
Struc:
1368 a.a.
1362 a.a.
Key:    PfamA domain  Secondary structure

 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     nucleic acid phosphodiester bond hydrolysis   3 terms 
  Biochemical function     hydrolase activity     9 terms  

 

 
DOI no: 10.1126/science.aad8282 Science 351:867-871 (2016)
PubMed id: 26841432  
 
 
Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage.
F.Jiang, D.W.Taylor, J.S.Chen, J.E.Kornfeld, K.Zhou, A.J.Thompson, E.Nogales, J.A.Doudna.
 
  ABSTRACT  
 
Bacterial adaptive immunity and genome engineering involving the CRISPR (clustered regularly interspaced short palindromic repeats)-associated (Cas) protein Cas9 begin with RNA-guided DNA unwinding to form an RNA-DNA hybrid and a displaced DNA strand inside the protein. The role of this R-loop structure in positioning each DNA strand for cleavage by the two Cas9 nuclease domains is unknown. We determine molecular structures of the catalytically active Streptococcus pyogenes Cas9 R-loop that show the displaced DNA strand located near the RuvC nuclease domain active site. These protein-DNA interactions, in turn, position the HNH nuclease domain adjacent to the target DNA strand cleavage site in a conformation essential for concerted DNA cutting. Cas9 bends the DNA helix by 30°, providing the structural distortion needed for R-loop formation.
 

 

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