Fires shape the biogeochemistry and functioning of many ecosystems, but fire frequencies are 1 changing across large areas of the globe. Frequent fires can change soil carbon (C) and nitrogen 2 (N) storage through both "top-down" pathways, by altering inputs through shifting plant 3 composition and biomass, and "bottom-up" ones, by altering losses through decomposition and 4 turnover of soil organic matter. However, the relative importance of these different pathways and 5 the degree to which they regulate ecosystem responses to decades of changing fire frequencies is 6 uncertain. Here, we sampled soils and plant communities in four North American and African 7 sites spanning tropical savanna, temperate coniferous savanna, temperate broadleaf savanna, and 8 temperate coniferous forest that each contained multiple plots repeatedly burned for 33-61 years 9 and nearby plots that were protected from fire over the same period. The sites varied markedly in 10 temperature, precipitation, species composition, fire history and soil chemistry; thus they 11 represent a broad test for the generality of fire impacts on biogeochemical cycling. For all four 12 sites, bulk soil C and N by were 25-180% higher in unburned vs. frequently burned plots, with 13 greater soil losses occurring in areas with greater declines in tree cover and biomass inputs into 14 soils. Fire reduced the activity of soil extracellular enzymes that hydrolyze labile C and N from 15 soil organic matter by two-to ten-fold, whereas tree cover was the predominant control on the 16 oxidation of recalcitrant C compounds. C-acquisition enzyme activity tended to decline with 17 decreasing soil N, suggesting that N losses may contribute to limited decomposition, buffering 18 systems against increased losses of soil C with fire. In conclusion, variability in how fire alters 19 soil C and N across ecosystems can be explained partly by fire-driven changes in tree cover and 20 biomass, but the slower turnover of organic matter we observed may offset some of the reduction 21 of C inputs from plants after fire. 22 Ecosystem responses to repeated burning 3 24 65 (Melillo et al. 1982)). Measuring the responses of extracellular enzymes provides insight into 66 these different factors because it disentangles the potential decomposition of different forms of 67 organic matter and the acquisition of C and N compounds separately. 68 Ecosystem responses to repeated burning 5 Here we evaluate the extent to which fire effects on soil C and N are driven by losses in 69 plant biomass inputs compared with changes in decomposition activity. Specifically, we 70 investigated (i) whether landscapes burned repeatedly have lower total soil C and N than those 71 protected from fire, including the degree to which this potential change is caused by direct 72 combustion compared with changes in tree cover and local enrichment (Pathway #1 in Figure 1); 73 and (ii) how fire changes soil C and N turnover, including decomposition of particular forms of 74 organic matter (Pathway #2 in Figure 1). ...