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PDBsum entry 4oo8

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protein dna_rna links
Hydrolase/DNA/RNA PDB id
4oo8
Jmol PyMol
Contents
Protein chains
1301 a.a.
1163 a.a.
DNA/RNA
Waters ×144
PDB id:
4oo8
Name: Hydrolase/DNA/RNA
Title: Crystal structure of streptococcus pyogenes cas9 in complex RNA and target DNA
Structure: Crispr-associated endonuclease cas9/csn1. Chain: a, d. Engineered: yes. Mutation: yes. RNA (97-mer). Chain: b, e. Engineered: yes. Other_details: single guide RNA. DNA (5'-
Source: Streptococcus pyogenes serotype m1. Organism_taxid: 301447. Gene: cas9. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes. Synthetic: yes
Resolution:
2.50Å     R-factor:   0.224     R-free:   0.254
Authors: H.Nishimasu,R.Ishitani,O.Nureki
Key ref: H.Nishimasu et al. (2014). Crystal structure of Cas9 in complex with guide RNA and target DNA. Cell, 156, 935-949. PubMed id: 24529477 DOI: 10.1016/j.cell.2014.02.001
Date:
31-Jan-14     Release date:   26-Feb-14    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
Q99ZW2  (Q99ZW2_STRP1) -  CRISPR-associated endonuclease Cas9/Csn1
Seq:
Struc:
 
Seq:
Struc:
 
Seq:
Struc:
1368 a.a.
1301 a.a.*
Protein chain
Pfam   ArchSchema ?
Q99ZW2  (Q99ZW2_STRP1) -  CRISPR-associated endonuclease Cas9/Csn1
Seq:
Struc:
 
Seq:
Struc:
 
Seq:
Struc:
1368 a.a.
1163 a.a.*
Key:    PfamA domain  Secondary structure
* PDB and UniProt seqs differ at 7 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: Chains A, D: E.C.3.1.-.-
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     nucleic acid phosphodiester bond hydrolysis   3 terms 
  Biochemical function     hydrolase activity     9 terms  

 

 
DOI no: 10.1016/j.cell.2014.02.001 Cell 156:935-949 (2014)
PubMed id: 24529477  
 
 
Crystal structure of Cas9 in complex with guide RNA and target DNA.
H.Nishimasu, F.A.Ran, P.D.Hsu, S.Konermann, S.I.Shehata, N.Dohmae, R.Ishitani, F.Zhang, O.Nureki.
 
  ABSTRACT  
 
The CRISPR-associated endonuclease Cas9 can be targeted to specific genomic loci by single guide RNAs (sgRNAs). Here, we report the crystal structure of Streptococcus pyogenes Cas9 in complex with sgRNA and its target DNA at 2.5 Å resolution. The structure revealed a bilobed architecture composed of target recognition and nuclease lobes, accommodating the sgRNA:DNA heteroduplex in a positively charged groove at their interface. Whereas the recognition lobe is essential for binding sgRNA and DNA, the nuclease lobe contains the HNH and RuvC nuclease domains, which are properly positioned for cleavage of the complementary and noncomplementary strands of the target DNA, respectively. The nuclease lobe also contains a carboxyl-terminal domain responsible for the interaction with the protospacer adjacent motif (PAM). This high-resolution structure and accompanying functional analyses have revealed the molecular mechanism of RNA-guided DNA targeting by Cas9, thus paving the way for the rational design of new, versatile genome-editing technologies.
 

 

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