Landscape change is an ongoing process for even the most established landscapes, especially in context to urban intensification and growth. As urbanization increases over the next century, supporting bird species’ populations within urbanizing areas remains an important conservation challenge. Fundamental elements of the biophysical structure of urban environments in which bird species likely respond include tree cover and human infrastructure. We broadly examine how tree cover and urban development structure bird species distributions along the urban-rural gradient across multiple spatial scales. We established a regional sampling design within the Oak Openings Region of northwestern, Ohio, USA, to survey bird species distributions across an extensive urbanization gradient. Through occupancy modeling, we obtained standardized effects of bird species response to local and landscape-scale predictors and found that landscape tree cover influenced the most species, followed by landscape impervious surface, local building density, and local tree cover. We found that responses varied according to habitat affiliation and migratory distance of individual bird species. Distributions of short-distance, edge habitat species located towards the rural end of the gradient were explained primarily by low levels of urbanization and potential vegetative and supplemental resources associated with these areas, while forest species distributions were primarily related to increasing landscape tree cover. Our findings accentuate the importance of scale relative to urbanization and help target where potential actions may arise to benefit bird diversity. Management will likely need to be implemented by municipal governments and agencies to promote tree cover at landscape scale, followed by residential land management education for private landowners. These approaches will be vital in sustaining biodiversity in urbanizing landscapes as urban growth expands over the next century.