The transition of land use function and its effects on ecosystem services is a key issue in eco-environmental protection and is the basis of territorial space governance and optimization. Previous studies have typically selected land use types to evaluate ecosystem service value (ESV) and have overlooked comprehensive characteristics of ecosystem services and the mutual feedback relationship between human social systems and the ecosystem. Taking the Three Gorges Reservoir Area, Hubei section (TGRA-HS) as a case study, we used a transition matrix, the revised ESV method, and an ecological contribution rate model to explore land use function transition (LUFT) and its effects on the change in ESV based on the production–living–ecological space (PLES) classification system. The results show that: (1) The transition of land use function based on PLES was the mapping of the evolution of the human–nature relationship in the spatial pattern, which reflected the evolution of the spatial pattern caused by human interference with the continuous development of society; (2) The evolution of PLES showed the characteristics of a reduction in production space (P-space), and an expansion in living space (L-space) and ecological space (E-space). The distribution pattern of PLES from 1990 to 2020 was basically the same, and the characteristics of structural transform reflected the characteristics of project construction in different phases; (3) The E-space contributed the most to the total ESV, and it has risen by CNY 13.06 × 10⁸. The transition of land use function caused by human construction projects impacts the spatiotemporal change in the regional ESV; (4) The change in ESV induced by LUFT revealed the whole dynamic process of the positive and negative effects of human construction projects on ecosystem services, and the two effects offset each other to keep the ESV relatively stable. The transition of E-space to P-space had the greatest impact on the reduction in ESV, whose contribution rate was 82.76%. The dynamic changes in land use function and ESV corresponding to the different stages of the Three Gorges Project’s (TGP) construction reveals the important driving effect of human activities on ecosystem services. It reminds us that humans should not forget to protect the eco-environment when obtaining services from the ecosystem.