3PKM

Crystal structure of Cas6 with its substrate RNA

Summary for 3PKM

• Entry DOI: 10.2210/pdb3pkm/pdb
• Descriptor: CRISPR-associated endoribonuclease Cas6, 5'-R(*AP*UP*UP*AP*CP*AP*AP*UP*AP*A)-3', 5'-R(P*UP*UP*AP*CP*AP*AP*UP*AP*A)-3' (3 entities in total)
• Functional Keywords: cas6, crispr, endonuclease, ferridoxin fold, crispr processing endonuclease, repeat rna, hydrolase-rna complex, hydrolase/rna
• Biological source: Pyrococcus furiosus; More
• Total number of polymer chains: 4
• Total formula weight: 68608.98

Authors

Wang, R.,Preamplume, G.,Li, H. (deposition date: 2010-11-11, release date: 2011-03-09, Last modification date: 2024-02-21)

Primary citation

Wang, R.,Preamplume, G.,Terns, M.P.,Terns, R.M.,Li, H.
Interaction of the Cas6 Riboendonuclease with CRISPR RNAs: Recognition and Cleavage.
Structure, 19:257-264, 2011

PubMed Abstract: The CRISPRs (Clustered Regularly Interspaced Short Palindromic Repeats) found in prokaryotic genomes confer small RNA-mediated protection against viruses and other invaders. CRISPR loci contain iterations of a short repeat sequence alternating with small segments of varying invader-derived sequences. Distinct families of CRISPR-associated Cas proteins function to cleave within the repeat sequence of CRISPR transcripts and produce the individual invader-targeting crRNAs. Here, we report the crystal structure of Pyrococcus furiosus Cas6 bound with a repeat RNA at 3.2 Å resolution. In contrast to other Cas families of endonucleases, Cas6 clasps nucleotides 2-9 of the repeat RNA using its two ferredoxin-like domains, and the enzyme-anchored 5' end tethers the distal cleavage site of the RNA between nucleotides 22 and 23 to the predicted enzyme active site on the opposite side of the ferrodoxin-like domains. Our findings suggest a wrap-around mechanism for CRISPR RNA recognition and cleavage by Cas6 and related processing endonucleases.

PubMed: 21300293
DOI: 10.1016/j.str.2010.11.014

Experimental method

X-RAY DIFFRACTION (3.103 Å)