Roads are widespread features of many landscapes that can negatively affect wildlife, most notably through animal-vehicle collisions. Roadside fencing has increasingly been installed to help eliminate this source of mortality. While fencing may reduce road mortality, other types of wildlife responses to this novel barrier are not well understood. Here, we examined the movement behavior, space use, and carapace temperatures of Mojave Desert Tortoises (Gopherus agassizii) as they interacted with a roadside fence and an unfenced road. Using GPS loggers, we tracked tortoise movements for two years at 15-min intervals. We found that carapace temperatures were greater near structures (fence or unfenced road) than away from structures; tortoises near the unfenced road had higher mean carapace temperatures, but tortoises along the fence experienced more extreme upper temperatures that approached the species' thermal limit. Movement speeds were also higher along the structures than away from them. Tortoise home range sizes decreased with proximity to the fence or road; fragmentation of home ranges and road-crossing avoidance may have contributed to smaller home ranges along the fenced and unfenced road, respectively. While tortoises crossed the road significantly less than expected by chance, they did so primarily in May and July and in areas with washes, indicating that placement of roadside fencing and animal underpasses could be optimized by targeting areas where roads intersect washes. Taken together, our results suggest that roadside fencing can affect behavior, space use, and thermal ecology of tortoises, which may require refinements to future conservation strategies involving roadside fencing.