Microorganisms in subglacial environments drive the chemical weathering of bedrock; however, the influence of bedrock mineralogy on the composition and activity of microbial assemblages in such environments is poorly understood. Here, using a combination of in situ mineral incubation and DNA fingerprinting techniques, we demonstrate that pyrite is the dominant mineralogical control on subglacial bacterial community structure and composition. In addition, we show that the abundance of Fe in the incubated minerals influences the development of mineral-associated biomass. The ubiquitous nature of pyrite in many common bedrock types and high SO42– concentrations in most glacial meltwaters suggest that pyrite may be a dominant lithogenic control on microbial communities in many subglacial systems. Mineral-based energy may therefore serve a fundamental role in sustaining subglacial microbial populations and enabling their persistence over glacial-interglacial time scales.