Climatic changes will not only result in higher overall temperature, but also in greater variability in weather conditions. Antarctic soils are subjected to extremely variable conditions in the form of frequent freeze–thaw cycles (FTCs), but the importance of alteration in FTC frequency, compared with increases in average temperature and indirect vegetation-mediated effects on soil microorganisms, is still unknown. We therefore designed two complementary microcosm experiments using undisturbed soil cores from Signy Island (60°43'S, 45°38'W) in the maritime Antarctic. The experiments consisted of soil core incubations with or without the overlying vegetation at four different temperatures and six different FTC regimes. We assessed bacterial and fungal density and community structure, as well as the density of several key genes in microbial nutrient cycles using a combination of RNA- and DNA-based molecular fingerprinting and quantitative PCR approaches in addition to enzymatic activity assays. Results showed that bacteria were more affected by warming than by changes in FTC frequency. In contrast, fungal community structure and abundance were mostly influenced by FTC frequency, as well as the presence of vegetation cover. The relative densities of several bacterial gene families involved in key steps of the N-cycle were affected by FTCs, while warming had little or no effect. The FTCs and incubation temperature also strongly influenced laccase enzymatic activity in soil. In total, our results suggest that, in addition to climatic warming, increased climatic variability may also have a profound impact on Antarctic microbial communities. Although these effects are difficult to detect with assays of total bacterial community structure, they do become manifest in the analysis of key functional gene densities.