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TitleMolecular mechanism for target recognition, dimerization, and activation of Pyrococcus furiosus Argonaute.
Journal, issue, pagesMol Cell, Vol. 84, Issue 4, Page 675-686.e4, Year 2024
Publish dateFeb 15, 2024
AuthorsLongyu Wang / Wanping Chen / Chendi Zhang / Xiaochen Xie / Fuyong Huang / Miaomiao Chen / Wuxiang Mao / Na Yu / Qiang Wei / Lixin Ma / Zhuang Li /
PubMed AbstractThe Argonaute nuclease from the thermophilic archaeon Pyrococcus furiosus (PfAgo) contributes to host defense and represents a promising biotechnology tool. Here, we report the structure of a PfAgo- ...The Argonaute nuclease from the thermophilic archaeon Pyrococcus furiosus (PfAgo) contributes to host defense and represents a promising biotechnology tool. Here, we report the structure of a PfAgo-guide DNA-target DNA ternary complex at the cleavage-compatible state. The ternary complex is predominantly dimerized, and the dimerization is solely mediated by PfAgo at PIWI-MID, PIWI-PIWI, and PAZ-N interfaces. Additionally, PfAgo accommodates a short 14-bp guide-target DNA duplex with a wedge-type N domain and specifically recognizes 5'-phosphorylated guide DNA. In contrast, the PfAgo-guide DNA binary complex is monomeric, and the engagement of target DNA with 14-bp complementarity induces sufficient dimerization and activation of PfAgo, accompanied by movement of PAZ and N domains. A closely related Argonaute from Thermococcus thioreducens adopts a similar dimerization configuration with an additional zinc finger formed at the dimerization interface. Dimerization of both Argonautes stabilizes the catalytic loops, highlighting the important role of Argonaute dimerization in the activation and target cleavage.
External linksMol Cell / PubMed:38295801
MethodsEM (single particle)
Resolution2.9 Å
Structure data

36489

8jpx

EMDB-36489, PDB-8jpx:
Cryo-EM structure of PfAgo-guide DNA-target DNA complex
Method: EM (single particle) / Resolution: 2.9 Å

37457

8wd8

EMDB-37457, PDB-8wd8:
Cryo-EM structure of TtdAgo-guide DNA-target DNA complex
Method: EM (single particle) / Resolution: 2.9 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-ZN:
Unknown entry

Source
  • pyrococcus furiosus (strain atcc 43587 / dsm 3638 / jcm 8422 / vc1) (archaea)
  • pyrococcus furiosus dsm 3638 (archaea)
  • thermus thermophilus (bacteria)
  • thermococcus thioreducens (archaea)
KeywordsGENE REGULATION / nuclease / DNA BINDING PROTEIN/DNA / DNA BINDING PROTEIN-DNA COMPLEX

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