6IV8
the selenomethionine(SeMet)-derived Cas13d binary complex
Summary for 6IV8
Entry DOI | 10.2210/pdb6iv8/pdb |
Descriptor | The selenomethionine (SeMet)-labeled Cas13d, RNA (51-MER), RNA (53-MER), ... (5 entities in total) |
Functional Keywords | crispr, cas13d, crrna, binary complex, rna binding protein-rna complex, rna binding protein/rna |
Biological source | uncultured Ruminococcus sp More |
Total number of polymer chains | 4 |
Total formula weight | 250691.09 |
Authors | Zhang, B.,Ye, Y.M.,Ye, W.W.,OuYang, S.Y. (deposition date: 2018-12-02, release date: 2019-06-19, Last modification date: 2024-10-23) |
Primary citation | Zhang, B.,Ye, Y.,Ye, W.,Perculija, V.,Jiang, H.,Chen, Y.,Li, Y.,Chen, J.,Lin, J.,Wang, S.,Chen, Q.,Han, Y.S.,Ouyang, S. Two HEPN domains dictate CRISPR RNA maturation and target cleavage in Cas13d. Nat Commun, 10:2544-2544, 2019 Cited by PubMed Abstract: Cas13d, the type VI-D CRISPR-Cas effector, is an RNA-guided ribonuclease that has been repurposed to edit RNA in a programmable manner. Here we report the detailed structural and functional analysis of the uncultured Ruminococcus sp. Cas13d (UrCas13d)-crRNA complex. Two hydrated Mg ions aid in stabilizing the conformation of the crRNA repeat region. Sequestration of divalent metal ions does not alter pre-crRNA processing, but abolishes target cleavage by UrCas13d. Notably, the pre-crRNA processing is executed by the HEPN-2 domain. Furthermore, both the structure and sequence of the nucleotides U(-8)-C(-1) within the repeat region are indispensable for target cleavage, and are specifically recognized by UrCas13d. Moreover, correct base pairings within two separate spacer regions (an internal and a 3'-end region) are essential for target cleavage. These findings provide a framework for the development of Cas13d into a tool for a wide range of applications. PubMed: 31186424DOI: 10.1038/s41467-019-10507-3 PDB entries with the same primary citation |
Experimental method | X-RAY DIFFRACTION (2.15 Å) |
Structure validation
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