Climate variability and human activity were regarded as two contributors to streamflow alteration. However, the contributions of the two factors were still unclear in Dongting Lake. Therefore, it was crucial to quantify the relative impact of climate variability and human activity on streamflow alteration. The time series (1961–2010) was divided into three periods, namely, natural period (1961–1980), change period I (1981–2002) and change period II (2003–2010). Sensitivity analysis based on Budyko-type equations was applied to reveal the contributions of climate variability and human activity in those two change periods, respectively. The results showed that during the change period I, climate variability was the main factor responsible for streamflow alteration in most parts of Dongting Lake, accounting for 60.07–67.27%. However, the impact of climate variability was slightly smaller than that of human activity in West Dongting Lake (the former accounting for 43.20% while the latter accounting for 56.80%). For the change period II, human activity was the dominate factor for streamflow alteration, accounting for 58.89–78.33%. The impact of climate variability gradually decreased while the impact of human activity gradually increased. Along with the intensification of the human activity, the impact of it became more dominant. The results could provide a reference for water resources planning and management decisions. Under the condition of uncontrollable climatic factor, effective measures should be put forward in controlling human activity, such as reservoir/dam operation, closed management of protected area and so on. Besides, it is essential to study the impact of climate variability on future water resources and water resource management under different climate change scenarios. Copyright © 2016 John Wiley & Sons, Ltd.