From DNA Groove Readability to Enzyme-Gated Opening: EcoRV as a First Test Case for Hydration Routing

This article proposes EcoRV–DNA–Mg²⁺ as a first enzyme-gated test case for hydration routing beyond residence time. It distinguishes local water presence from functional hydration routing and introduces routed-hydration observables for testing whether a hydration-ion configuration remains oriented, bridge-connected, reconfigurable, and compatible with a transition-supporting window during EcoRV pre-cleavage opening. The manuscript is a theoretical and computational proof-of-principle proposal. It does not report completed molecular dynamics datasets. Instead, it defines a falsifiable MD-only protocol, local analysis zones, routing-versus-residence observables, sensitivity controls, and future comparator systems for testing whether hydration routing provides functional information beyond residence time in enzyme-gated DNA opening.