Atmospheric deposition of reactive nitrogen (Nr) can impact the soil microbial community structure and function and thus ecosystem processing and export of nutrients. Ecosystem response to atmospheric inputs of nitrogen depends on several factors, including elevational climate conditions (freeze/thaw cycles, precipitation), geology, soil and vegetation type, N speciation and microbial community structure. The focus of this study was to evaluate how spatial and seasonal variations in N deposition affect the ability of soil microbial communities to process N along an elevational gradient (1700 to 3015 meters) from plains to subalpine ecosystems west of Boulder, Colorado. In conjunction with a study measuring the variability of wet-plus-dry Nr Deposition along this gradient using ion exchange resin (IER) columns (Heindel et al., 2022), soil samples (0–10 cm depth) were collected seasonally at 3 locations and once at a 4th location. To assess the role that the soil microbial community plays in processing nitrogen, soils were measured for nitrification potential, net N mineralization rates, microbial community structure, and N-cycling gene abundance. Additionally, soils were characterized for total carbon and nitrogen content, pH, water and potassium chloride extractable dissolved organic carbon and nitrogen species, ammonium sorption potential, and bulk density. Atmospheric deposition of reactive nitrogen (Nr) can impact the soil microbial community structure and function and thus ecosystem processing and export of nutrients. Ecosystem response to atmospheric inputs of nitrogen depends on several factors, including elevational climate conditions (freeze/thaw cycles, precipitation), geology, soil and vegetation type, N speciation and microbial community structure. The focus of this study was to evaluate how spatial and seasonal variations in N deposition affect the ability of soil microbial communities to process N along an elevational gradient (1700 to 3015 meters) from plains to subalpine ecosystems west of Boulder, Colorado. In conjunction with a study measuring the variability of wet-plus-dry Nr Deposition along this gradient using ion exchange resin (IER) columns (Heindel et al., 2022), soil samples (0–10 cm depth) were collected seasonally at 3 locations and once at a 4th location. To assess the role that the soil microbial community plays in processing nitrogen, soils were measured for nitrification potential, net N mineralization rates, microbial community structure, and N-cycling gene abundance. Additionally, soils were characterized for total carbon and nitrogen content, pH, water and potassium chloride extractable dissolved organic carbon and nitrogen species, ammonium sorption potential, and bulk density. Site information, soil characterization details, N-cycling nutrient data, and molecular results are presented in this data release as comma-separated values (.csv) formatted tables. A “Site Details” file provides general location information for the four study sites. Data files are grouped in zipped file folders under 2 categories – 1. Nutrient Data and 2. Microbial Data. Two data dictionary tables, Nutrient Data Descriptions and Microbial Data Descriptions, are located in the zipped folders and describe the data found in the .csv files. Method details and references are located in the .xml metadata file “N Dep Soil Activity.”