9CEW

Spizellomyces punctatus Fanzor (SpuFz) State 3

Summary for 9CEW

• Entry DOI: 10.2210/pdb9cew/pdb
• EMDB information: 45521
• Descriptor: DNA (5'-D(P*CP*GP*GP*TP*AP*CP*CP*CP*GP*GP*GP*CP*AP*TP*A)-3'), Maltose/maltodextrin-binding periplasmic protein,Spizellomyces punctatus Fanzor 1, DNA (29-MER), ... (8 entities in total)
• Functional Keywords: fanzor, eukaryotic, rna-guided, nuclease, gene editing, rna binding protein-rna-dna complex, rna binding protein/rna/dna
• Biological source: Escherichia coli K-12; More
• Total number of polymer chains: 6
• Total formula weight: 196257.79

Authors

Xu, P.,Saito, M.,Zhang, F. (deposition date: 2024-06-27, release date: 2024-09-11, Last modification date: 2024-10-02)

Primary citation

Xu, P.,Saito, M.,Faure, G.,Maguire, S.,Chau-Duy-Tam Vo, S.,Wilkinson, M.E.,Kuang, H.,Wang, B.,Rice, W.J.,Macrae, R.K.,Zhang, F.
Structural insights into the diversity and DNA cleavage mechanism of Fanzor.
Cell, 187:5238-5252.e20, 2024

PubMed Abstract: Fanzor (Fz) is an ωRNA-guided endonuclease extensively found throughout the eukaryotic domain with unique gene editing potential. Here, we describe the structures of Fzs from three different organisms. We find that Fzs share a common ωRNA interaction interface, regardless of the length of the ωRNA, which varies considerably across species. The analysis also reveals Fz's mode of DNA recognition and unwinding capabilities as well as the presence of a non-canonical catalytic site. The structures demonstrate how protein conformations of Fz shift to allow the binding of double-stranded DNA to the active site within the R-loop. Mechanistically, examination of structures in different states shows that the conformation of the lid loop on the RuvC domain is controlled by the formation of the guide/DNA heteroduplex, regulating the activation of nuclease and DNA double-stranded displacement at the single cleavage site. Our findings clarify the mechanism of Fz, establishing a foundation for engineering efforts.

PubMed: 39208796
DOI: 10.1016/j.cell.2024.07.050

Experimental method

ELECTRON MICROSCOPY (2.88 Å)