ABSTRACT Roadkill, the result of collisions between vehicles and wildlife, is becoming one of the most important conservation issues. This is especially evident in regions such as the Cerrado savannas which are under strong land use conversion, forcing animals to move more through the landscape and thus increasing exposure to roadkill. Herein, we assess the temporal and spatial patterns of roadkill of the striped hog-nosed skunk (Conepatus semistriatus) as well as its interaction with surrounding land use in the Brazilian Cerrado. To do so, we sampled BR-050, a nationally-important highway, for roadkilled C. semistriatus between 2015 and 2020 and used location and date to identify temporal and spatial patterns of roadkill. Furthermore, we assessed whether different land uses affected wildlife-vehicle collisions using a random forest model. We recorded 672 roadkilled individuals of C. semistriatus, and detected both monthly and yearly temporal hotspots as well as spatial hotspots. These hotspots were significantly surrounded by soy, which along with savannas and crops were the most impactful land uses. Since conservation practitioners need to know when and where there are higher risks of wildlife-vehicle collision, our results are essential for the conservation of C. semistriatus in the Cerrado by incorporating land use into roadkill.