5F9R
Crystal structure of catalytically-active Streptococcus pyogenes CRISPR-Cas9 in complex with single-guided RNA and double-stranded DNA primed for target DNA cleavage
Summary for 5F9R
| Entry DOI | 10.2210/pdb5f9r/pdb |
| Descriptor | CRISPR-associated endonuclease Cas9/Csn1, RNA (116-MER), DNA (30-MER), ... (5 entities in total) |
| Functional Keywords | crispr, cas9, r-loop, genome engineering, hydrolase-dna-rna complex, hydrolase/dna/rna |
| Biological source | Streptococcus pyogenes serotype M1 More |
| Total number of polymer chains | 4 |
| Total formula weight | 215728.56 |
| Authors | Jiang, F.,Doudna, J.A. (deposition date: 2015-12-10, release date: 2016-01-27, Last modification date: 2024-03-06) |
| Primary citation | Jiang, F.,Taylor, D.W.,Chen, J.S.,Kornfeld, J.E.,Zhou, K.,Thompson, A.J.,Nogales, E.,Doudna, J.A. Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage. Science, 351:867-871, 2016 Cited by PubMed 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. PubMed: 26841432DOI: 10.1126/science.aad8282 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (3.4 Å) |
Structure validation
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