Running during sports and for physical activity often requires changes in velocity through acceleration and deceleration. While it is clear that lower extremity biomechanics vary during these accelerations and decelerations, the work requirements of the individual joints are not well understood. The purpose of this investigation was to measure the sagittal plane mechanical work of the individual lower extremity joints during acceleration, deceleration, and steady-state running. Ten runners were compared during acceleration, deceleration, and steady-state running using three-dimensional kinematics and kinetics measures. Total positive and negative joint work, and relative joint contributions to total work were compared between conditions. Total positive work progressively increased from deceleration to acceleration. This was due to greater ankle joint work during acceleration. While there was no significant change in total negative work during deceleration, there was a greater relative contribution of the knee to total negative work with a subsequent lower relative ankle negative work. Each lower extremity joint exhibits distinct functional roles in acceleration compared with deceleration during level running. Deceleration is dominated by greater contributions of the knee to negative work while acceleration is associated with a greater ankle contribution to positive work.