Hydrogen embrittlement (HE) causes engineering alloys to fracture unexpectedly, often at considerable economic or environmental cost. Inaccurate predictions of component lifetimes arise from inadequate understanding of how alloy microstructure affects HE. Here we investigate hydrogen-assisted fracture of a Ni-base superalloy and identify coherent twin boundaries (CTBs) as the microstructural features most susceptible to crack initiation. This is a surprising result considering the renowned beneficial effect of CTBs on mechanical strength and corrosion resistance of many engineering alloys. Remarkably, we also find that CTBs are resistant to crack propagation, implying that hydrogen-assisted crack initiation and propagation are governed by distinct physical mechanisms in Ni-base alloys. This finding motivates a re-evaluation of current lifetime models in light of the dual role of CTBs. It also indicates new paths to designing materials with HE-resistant microstructures.