On a global scale, human activity has increased the atmospheric input of NO3- to many terrestrial ecosystems. Anthropogenic NO3- may be a potent modifier of ecosystem function, especially in temperate forests that are sometimes N limited. However, the impact of chronic N deposition on soil microorganisms is still poorly understood. Nitrate entering Lake States forests is rapidly assimilated by the microbial community and it is subsequently released as NH4+. Because high levels of NH4+ inhibit the activity of lignin-degrading soil fungi, we reasoned that chronic N additions could alter the composition and function of heterotrophic microbial communities in soil, and hence the ecosystem-level processes they mediate. We tested our hypothesis in four northern hardwood ecosystems in northern Michigan, which received experimental N additions (30 kg NO3-/ha/yr)during the past 8 yr. We quantified microbial community function by measuring the activity of extracellular enzymes involved in plant litter degradation and described microbial community composition using phospholipid fatty acid (PFLA) analysis. Chronic N additions significantly suppressed B-glucosidase activity by 24% in mineral soil and suppressed phenol oxidase activity by 35% in surface litter. We found no evidence that chronic N additions altered microbial community composition; NO3- addition did not alter the relative abundance of bacterial, actinomycetal, fungal, or protozoan PFLAs. However, NO3- additions significantly reduced microbial biomass by 18% relative to the control treatment. Results indicate that N additions broadly suppressed all microbial groups, not just the activity and abundance of lignin-degrading fungi.