Understanding how nitrogen transport across the landscape varies with landscape characteristics is important for developing sound nitrogen management policies. We used a spatially referenced regression analysis (SPARROW) to examine landscape characteristics influencing delivery of nitrogen from sources in a watershed to stream channels. Modelled landscape delivery ratio varies widely (by a factor of 4) among watersheds in the southeastern United States—higher in the western part (Tennessee, Alabama, and Mississippi) than in the eastern part, and the average value for the region is lower compared to other parts of the nation. When we model landscape delivery ratio as a continuous function of local-scale landscape characteristics, we estimate a spatial pattern that varies as a function of soil and climate characteristics but exhibits spatial structure in residuals (observed load minus predicted load). The spatial pattern of modelled landscape delivery ratio and the spatial pattern of residuals coincide spatially with Level III ecoregions and also with hydrologic landscape regions. Subsequent incorporation into the model of these frameworks as regional scale variables improves estimation of landscape delivery ratio, evidenced by reduced spatial bias in residuals, and suggests that cross-scale processes affect nitrogen attenuation on the landscape. The model-fitted coefficient values are logically consistent with the hypothesis that broad-scale classifications of hydrologic response help to explain differential rates of nitrogen attenuation, controlling for local-scale landscape characteristics. Negative model coefficients for hydrologic landscape regions where the primary flow path is shallow ground water suggest that a lower fraction of nitrogen mass will be delivered to streams; this relation is reversed for regions where the primary flow path is overland flow.