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PDBsum entry 4zt9

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protein dna_rna links
Hydrolase/RNA PDB id
4zt9
Jmol PyMol
Contents
Protein chains
1335 a.a.
1160 a.a.
DNA/RNA
PDB id:
4zt9
Name: Hydrolase/RNA
Title: Nuclease-inactive streptococcus pyogenes cas9 (d10a/h840a, d complex with single-guide RNA at 3.1 angstrom resolution
Structure: Crispr-associated endonuclease cas9. Chain: a, c. Synonym: cas9, crispr-cas9. Engineered: yes. Mutation: yes. Single-guide RNA. Chain: b, d. Engineered: yes
Source: Streptococcus pyogenes. Organism_taxid: 1314. Gene: csn1, cas9, ers445054_00848, mtb314_0777. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008. Expressed in: escherichia coli. Expression_system_taxid: 562
Resolution:
3.10Å     R-factor:   0.245     R-free:   0.284
Authors: F.Jiang,J.A.Doudna
Key ref: F.Jiang et al. (2015). STRUCTURAL BIOLOGY. A Cas9-guide RNA complex preorganized for target DNA recognition. Science, 348, 1477-1481. PubMed id: 26113724 DOI: 10.1126/science.aab1452
Date:
14-May-15     Release date:   08-Jul-15    
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.
1335 a.a.*
Protein chain
Pfam   ArchSchema ?
Q99ZW2  (Q99ZW2_STRP1) -  CRISPR-associated endonuclease Cas9/Csn1
Seq:
Struc:
 
Seq:
Struc:
 
Seq:
Struc:
1368 a.a.
1160 a.a.*
Key:    PfamA domain  Secondary structure
* PDB and UniProt seqs differ at 6 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: Chains A, C: 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: 10.1126/science.aab1452 Science 348:1477-1481 (2015)
PubMed id: 26113724  
 
 
STRUCTURAL BIOLOGY. A Cas9-guide RNA complex preorganized for target DNA recognition.
F.Jiang, K.Zhou, L.Ma, S.Gressel, J.A.Doudna.
 
  ABSTRACT  
 
Bacterial adaptive immunity uses CRISPR (clustered regularly interspaced short palindromic repeats)-associated (Cas) proteins together with CRISPR transcripts for foreign DNA degradation. In type II CRISPR-Cas systems, activation of Cas9 endonuclease for DNA recognition upon guide RNA binding occurs by an unknown mechanism. Crystal structures of Cas9 bound to single-guide RNA reveal a conformation distinct from both the apo and DNA-bound states, in which the 10-nucleotide RNA "seed" sequence required for initial DNA interrogation is preordered in an A-form conformation. This segment of the guide RNA is essential for Cas9 to form a DNA recognition-competent structure that is poised to engage double-stranded DNA target sequences. We construe this as convergent evolution of a "seed" mechanism reminiscent of that used by Argonaute proteins during RNA interference in eukaryotes.
 

 

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