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TitleStructural basis for the type I-F Cas8-HNH system.
Journal, issue, pagesEMBO J, Year 2024
Publish dateSep 9, 2024
AuthorsXuzichao Li / Yanan Liu / Jie Han / Lingling Zhang / Zhikun Liu / Lin Wang / Shuqin Zhang / Qian Zhang / Pengyu Fu / Hang Yin / Hongtao Zhu / Heng Zhang /
PubMed AbstractThe Cas3 nuclease is utilized by canonical type I CRISPR-Cas systems for processive target DNA degradation, while a newly identified type I-F CRISPR variant employs an HNH nuclease domain from the ...The Cas3 nuclease is utilized by canonical type I CRISPR-Cas systems for processive target DNA degradation, while a newly identified type I-F CRISPR variant employs an HNH nuclease domain from the natural fusion Cas8-HNH protein for precise target cleavage both in vitro and in human cells. Here, we report multiple cryo-electron microscopy structures of the type I-F Cas8-HNH system at different functional states. The Cas8-HNH Cascade complex adopts an overall G-shaped architecture, with the HNH domain occupying the C-terminal helical bundle domain (HB) of the Cas8 protein in canonical type I systems. The Linker region connecting Cas8-NTD and HNH domains adopts a rigid conformation and interacts with the Cas7.6 subunit, enabling the HNH domain to be in a functional position. The full R-loop formation displaces the HNH domain away from the Cas6 subunit, thus activating the target DNA cleavage. Importantly, our results demonstrate that precise target cleavage is dictated by a C-terminal helix of the HNH domain. Together, our work not only delineates the structural basis for target recognition and activation of the type I-F Cas8-HNH system, but also guides further developments leveraging this system for precise DNA editing.
External linksEMBO J / PubMed:39251884
MethodsEM (single particle)
Resolution2.51 - 3.6 Å
Structure data

39706

8z0k

EMDB-39706, PDB-8z0k:
Cryo-EM structure of Cas8-HNH system at full R-loop state
Method: EM (single particle) / Resolution: 2.51 Å

39707

8z0l

EMDB-39707, PDB-8z0l:
Cryo-EM structure of Cas8-HNH system at partial R-loop state
Method: EM (single particle) / Resolution: 2.57 Å

60017

8zdy

EMDB-60017, PDB-8zdy:
Cryo-EM structure of Cas8-HNH system at target free state
Method: EM (single particle) / Resolution: 3.6 Å

60279

8znr

EMDB-60279, PDB-8znr:
Cryo-EM structure of Cas8-HNH system at ssDNA-bound state
Method: EM (single particle) / Resolution: 2.9 Å

Source
  • selenomonas sp. (bacteria)
KeywordsANTIVIRAL PROTEIN/DNA/RNA / a protein complex / ANTIVIRAL PROTEIN-DNA-RNA complex / ANTIVIRAL PROTEIN

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