Streams and rivers in urban watersheds are predicted to export more bioreactive, autochthonous dissolved organic matter (DOM) relative to forested watersheds. However, the spatial and temporal variations of DOM quality in forested urban watersheds remain uncertain, and their relationships with socioeconomic conditions, biological characteristics, and the built environment are understudied. We measured optical properties of fluorescent DOM (FDOM) in 93 streams spanning a gradient of land-use and land cover during four seasons in metropolitan Atlanta, Georgia, USA. Streamwater FDOM was dominated by humic substances from anthropogenic (41%) and terrestrial origin (41.5%). Impervious surface cover was the strongest predictor, which was positively correlated with anthropogenically- and autochthonously-derived FDOM. Overwater canopy cover was positively associated with autochthonous FDOM, and housing age increased diagenetic FDOM. FDOM was more proteinaceous during low-flow conditions (fall, winter), and more allochthonous humic-like FDOM was detected during periods of higher flows (spring, summer). Interestingly, wastewater-related FDOM proxies were highest during low flows, suggesting that sewer exfiltration is a pervasive source and is diluted by other inputs during high flows. Overall, seasonal patterns in FDOM quality were associated with changes in hydrology, and FDOM was primarily humic throughout the year, a pattern likely driven by ubiquitous forest canopy cover. Our results highlight the importance of urban forests in mediating aquatic carbon cycling and provide a template for future studies that integrate sociodemographic and infrastructure information into studies of watershed biogeochemistry, especially in regions undergoing rapid, intense, and localized urban development.