Streams are potential hotspots for retention and removal of NO₃⁻, and understanding the mechanisms that enhance NO₃⁻ reactivity in stream systems is critical for predicting and preventing eutrophication. Both dissolved organic C (DOC) and dissolved O₂ (DO) influence NO₃⁻ removal processes. Assessing the individual impacts of NO₃⁻, DO, and DOC concentrations on stream NO₃⁻ removal is difficult because these factors covary and are coupled through the C and N cycles. We used an experimental approach to quantify the influences of NO₃⁻, DOC, and DO on NO₃⁻ transport in headwater streams of the Ipswich and Parker River watersheds (Massachusetts, USA) with contrasting levels of DOC and DO. In a 1^st set of experiments, we added NO₃⁻ to address how uptake kinetics differed between a low-DO/high-DOC stream (Cedar Swamp Creek) and a high-DO/low-DOC stream (Cart Creek). In a 2^nd set of experiments, we manipulated, for the first time at the reach scale, both DO and DOC in a factorial experiment. DO was added to the low-DO stream by injecting O₂ and was removed from the high-DO stream by adding sodium sulfite. DOC was added both alone and in combination with the DO manipulations. NO₃⁻ concentration was an important control of NO₃⁻ uptake velocity in our study streams, consistent with previous findings. The results of the DOC and DO manipulations suggested that DO determines whether a stream has net NO₃⁻ uptake or production and that the presence of DOC magnifies the DO response processes. Addition of DOC by itself did not lead to increased NO₃⁻ uptake. In addition, we observed organic matter priming effects, wherein the addition of labile organic matter resulted in accelerated metabolism of naturally occurring DOC in the water column. Priming effects have not been reported previously in stream systems. Results from our experiments suggest that NO₃⁻ uptake in streams might arise from complex interactions among DOC, DO, and NO₃⁻, and ultimately, from the influence of DO on dominant stream processes.