Rapidly expanding road networks have been a key driver of the fragmentation and isolation of many wildlife species, and are a source of significant mortality due to collisions with vehicles. But not all animals are affected equally by transportation infrastructure, and in most cases little is known about the population-scale consequences of roads for wildlife. Even less information is available to characterize species’ behavioral responses to roads. Although research shows that maned wolves (Chrysocyon brachyurus) in Brazil are experiencing considerable fragmentation and road mortality, the degree to which these disturbances are impacting the species’ viability is unknown. The goal of this study was to analyze the potential effects of roads on maned wolf population size and structure. We used a simulation model to evaluate the population-scale consequences of individual maned wolf interactions with roads, which can result in road crossing, avoidance, or mortality due to a collision with a vehicle. We also forecasted where in Brazil these impacts might be most significant. Our model incorporated species demographic and movement parameters, plus habitat quality and a map of the road network. We found that even moderate rates of road mortality led to severe declines in population size, and that four specific locations accounted for a disproportionate fraction of roadkill events. Our approach will be generally useful for evaluating the relative importance of road effects on species conservation in many ecological systems, for prioritizing data collection efforts, and for informing conservation policies and mitigation strategies.