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-Structure paper
Title | Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage. |
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Journal, issue, pages | Science, Vol. 351, Issue 6275, Page 867-871, Year 2016 |
Publish date | Feb 19, 2016 |
Authors | Fuguo Jiang / David W Taylor / Janice S Chen / Jack E Kornfeld / Kaihong Zhou / Aubri J Thompson / Eva Nogales / Jennifer A Doudna / |
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 ...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. |
External links | Science / PubMed:26841432 / PubMed Central |
Methods | EM (single particle) / X-ray diffraction |
Resolution | 3.4 - 6.0 Å |
Structure data | EMDB-3276: EMDB-3277: PDB-5f9r: |
Chemicals | ChemComp-SO4: |
Source |
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Keywords | HYDROLASE/DNA/RNA / CRISPR / Cas9 / R-loop / genome engineering / HYDROLASE-DNA-RNA complex |