The effects of stream size and flow regime on spatial and temporal variability of stream fish distribution, abundance, and diversity patterns were investigated. Assemblage variability and species richness were each significantly associated with a complex environmental gradient contrasting smaller, hydrologically variable stream localities with larger localities characterized by more stable flow regimes. Assemblages showing the least variability were the most species-rich and occurred in relatively large, stable environments. Theory suggests that species richness can be an important determinant of assemblage variability. Although this appears to be true in our system, we suggest that spatial and temporal heterogeneity in the environment largely determines both assemblage richness and variability, providing a more parsimonious explanation for the diversity–variability correlation. Changes in species richness of local assemblages across time were coordinated across the landscape, and assemblages formed spatially and temporally nested subset patterns. These results suggest an important link between local community dynamics and community-wide occurrence. At the species level, mean local persistence was significantly associated with regional occurrence. Thus, the more widespread a species was, the greater its local persistence. Our results illustrate how the integrity of local stream fish assemblages is dependent on local environmental conditions, regional patterns of species distribution, and landscape continuity.