OBJECTIVE Successful weight management relies heavily on eating and exercise behaviors. However, little is known about the association between both on a psychosocial level. This study examined the relationship between exercise and eating regulation by exploring the mediating effects of negative body image investment and depressive mood, and their stability through time. METHODS Analyses were conducted at two different moments (12 and 36 months), involving a sample of 221 overweight/obese women (age: 37.6 ± 7 years; BMI: 31.6 ± 4.1 kg/m(2)) that participated in a behavioral weight control intervention. Bivariate correlations and mediation analyses using Preacher & Hayes resampling procedures were conducted. RESULTS At 12 months, negative body image investment was the only significant mediator of the exercise-eating relationship. This variable explained larger portions of the indirect effects of structured rather than lifestyle exercise on eating. At 36 months, negative investment and to a lesser extent depressive mood partially explained the exercise-eating association. CONCLUSIONS Our findings suggest that, besides physiological effects of exercise, psychological mechanisms related to body image and mood also explain the role of physical activity as a 'gateway behavior' for improved eating regulation in overweight women. These effects appear to be stable and may help understand the key role of exercise in long-term weight management.