A Cryptic Binding Pocket Regulates the Metal-Dependent Activity of Cas9
Ahsan, M.; Saha, A.; Ramos, D.; Strohkendl, I.; Knight, A. L.; Skeens, E.; Lisi, G. P.; Taylor, D. W.; Palermo, G.
Show abstract
Cas9 is a metal-dependent nuclease that has revolutionized gene editing across diverse cells and organisms exhibiting varying ion uptake, metabolism, and concentrations. However, how divalent metals impact its catalytic function, and consequently its editing efficiency in different cells, remains unclear. Here, extensive molecular simulations, Markov State Models, biochemical and NMR experiments, demonstrate that divalent metals - Mg2+, Ca2+, and Co2+ - promote activation of the catalytic HNH domain by binding within a dynamically forming divalent metal binding pocket (DBP) at the HNH-RuvC interface. Mutations in DBP residues disrupt HNH activation and impair the coupled catalytic activity of both nucleases, identifying this cryptic DBP as a key regulator of Cas9s metal-dependent activity. The ionic strength thereby promotes Cas9s conformational activation, while its catalytic activity is metal-specific. These findings are critical to improving the metal-dependent function of Cas9 and its use for genome editing in different cells and organisms.
Matching journals
The top 3 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Identifying Sequence Perturbations to an Intrinsically Disordered Protein that Determine Its Phase Separation Behavior 95%
- DNA Binding Induces a cis to trans Switch in Cre Recombinase to Enable Intasome Assembly 95%
- The conformational landscape of fold-switcher KaiB is tuned to the circadian rhythm timescale 95%
Similar papers in this journal
- Structural insights into the opening mechanism of C1C2 channelrhodopsin 97%
- Water-triggered, irreversible conformational change of SARS-CoV-2 main protease on passing from the solid state to aqueous solution 96%
- Chemical mechanism of allosteric and asymmetric dark reversion in a bacterial phytochrome uncovered by cryo-EM. 96%
Similar papers in this journal
- Determinants of sugar-induced influx in the mammalian fructose transporter GLUT5 96%
- Allosteric coupling asymmetry mediates paradoxical activation of BRAF by type II inhibitors 96%
- Observing one-divalent-metal-ion dependent and histidine-promoted His-Me family I-PpoI nuclease catalysis in crystallo 96%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.