Soil microbes comprise a large portion of the genetic diversity on Earth and influence a large number of important ecosystem processes. Increasing atmospheric nitrogen (N) deposition represents a major global change driver; however, it is still debated whether the impacts of N deposition on soil microbial biomass and respiration are ecosystem-type dependent. Moreover, the extent of N deposition impacts on microbial composition remains unclear. Here we conduct a global meta-analysis using 1408 paired observations from 151 studies to evaluate the responses of soil microbial biomass, composition, and function to N addition. We show that nitrogen addition reduced total microbial biomass, bacterial biomass, fungal biomass, biomass carbon, and microbial respiration. Importantly, these negative effects increased with N application rate and experimental duration. Nitrogen addition reduced the fungi to bacteria ratio and the relative abundances of arbuscular mycorrhizal fungi and gram-negative bacteria and increased gram-positive bacteria. Our structural equation modeling showed that the negative effects of N application on soil microbial abundance and composition led to reduced microbial respiration. The effects of N addition were consistent across global terrestrial ecosystems. Our results suggest that atmospheric N deposition negatively affects soil microbial growth, composition, and function across all terrestrial ecosystems, with more pronounced effects with increasing N deposition rate and duration.