The ability of an organism to disperse to suitable habitats, especially in modified and fragmented systems, determines individual fitness and overall population viability. The bluehead sucker (Catostomus discobolus), flannelmouth sucker (Catostomus latipinnis), and roundtail chub (Gila robusta) are three species native to the upper Colorado River Basin that now occupy only 50% of their historic range. Despite these distributional declines, populations of all three species are present in the San Rafael River, a highly regulated tributary of the Green River, Utah, providing an opportunity for research. Our goal was to determine the timing and extent of movement, habitat preferences, and limiting factors, ultimately to guide effective management and recovery of these three species. In 2007-2008, we sampled fish from 25 systematically selected, 300-m reaches in the lower 64 km of the San Rafael River, spaced to capture the range of species, life-stages, and habitat conditions present. We implanted all target species with a passive integrated transponder (PIT) tag, installed a passive PIT tag antennae, and measured key habitat parameters throughout each reach and at the site of native fish capture. We used random forest modeling to identify and rank the most important abiotic and biotic predictor variables, and reveal potential limiting factors in the San Rafael River. While flannelmouth sucker were relatively evenly distributed within our study area, highest densities of roundtail chub and bluehead sucker occurred in isolated, upstream reaches characterized by complex habitat. In addition, our movement and length-frequency data indicate downstream drift of age-0 roundtail chub, and active upstream movement of adult flannelmouth sucker, both from source populations, providing the lower San Rafael River with colonists. Our random forest analysis highlights the importance of pools, riffles, and distance-to-source populations, suggesting that bluehead sucker and roundtail chub are habitat limited in the lower San Rafael River. These results suggest management efforts should focus on diversifying habitat, maintaining in-stream flow, and removing barriers to movement.