Nature Communications· 2026Q1
A cryptic binding pocket regulates the metal-dependent activity of Cas9
- 3citations
- Q1SCImago
- 2026year
Short summary
A cryptic, dynamically formed divalent metal-binding pocket (DBP) at the HNH–RuvC interface regulates Cas9 nuclease activity, as revealed by simulations and NMR.
AI-generated from the title and abstract; the full text is not read.
Key points
- A cryptic divalent metal-binding pocket (DBP) dynamically forms at the HNH–RuvC interface of Cas9.
- This DBP is essential for metal ion-dependent activation of the HNH nuclease domain.
- NMR measurements confirm metal-dependent structural changes in the HNH–L2 region, supporting the DBP's existence.
- Mutations targeting the DBP disrupt HNH activation and impair Cas9's coupled catalytic activity.
AI-generated from the title and abstract; the full text is not read.
Abstract
Abstract Cas9 is a metal-dependent nuclease used for genome editing across diverse cells and organisms exhibiting distinct ionic environments, yet how metal ions regulate its catalytic function, and consequently its editing efficiency remains unclear. Here, molecular simulations, Markov state models, and mixed quantum–classical approaches reveal that divalent metals promote activation of the catalytic HNH domain through formation of a divalent metal-binding pocket (DBP) at the HNH–RuvC interface. This cryptic pocket emerges dynamically during HNH activation and is supported by NMR measurements revealing metal-dependent structural perturbations within the HNH–L2 region. Mutations targeting the DBP disrupt HNH activation and impair the coupled catalytic activity of both nucleases, identifying the pocket as a key regulator of Cas9’s metal-dependent activity. Together with quantum–classical simulations revealing distinct catalytic behaviours among divalent metals, these findings uncover an ion-dependent regulatory mechanism in Cas9 with insights for genome editing across diverse biological environments.
The authors' abstract, as published at the source. Nature Communications, 2026 · DOI ↗
Continue with a free account
Ask the paper: 3 free questions a day about this paper; save it, get its citation, new summaries every day for your field. Takeaways are Premium.
Continue free on the webSign in with Google or Apple; no card needed. You come back to this paper.
On your phone:
Field: Molecular Biology
Molecular BiologyBiochemistry, Genetics and Molecular Biology