Global climate models predict significant changes to the rainfall regimes of the grassland biome, where C cycling is particularly sensitive to the amount and timing of precipitation. We explored the effects of both natural interannual rainfall variability and experimental rainfall additions on net C storage and loss in annual grasslands. Soil respiration and net primary productivity (NPP) were measured in treatment and control plots over four growing seasons (water years, or WYs) that varied in wet-season length and the quantity of rainfall. In treatment plots, we increased total rainfall by 50% above ambient levels and simulated one early- and one late-season storm. The early- and late-season rain events significantly increased soil respiration for 2–4 weeks after wetting, while augmentation of wet-season rainfall had no significant effect. Interannual variability in precipitation had large and significant effects on C cycling. We observed a significant positive relationship between annual rainfall and aboveground NPP across the study (P=0.01, r2=0.69). Changes in the seasonal timing of rainfall significantly affected soil respiration. Abundant rainfall late in the wet season in WY 2004, a year with average total rainfall, led to greater net ecosystem C losses due to a ∼50% increase in soil respiration relative to other years. Our results suggest that C cycling in annual grasslands will be less sensitive to changes in rainfall quantity and more affected by altered seasonal timing of rainfall, with a longer or later wet season resulting in significant C losses from annual grasslands.