Background: Previous research suggested that modifiable health factors such as physical activity, sedentary behavior, and sleep (i.e., 24-hour time-use behaviors) were potential intervention targets to improve antenatal depressive symptoms. However, most previous studies examine one behavior at a time, ignoring their co-dependence. The aim of this study was to examine the association between 24-hour time-use behaviors and depressive symptoms during early pregnancy. Methods: We conducted a secondary analysis using baseline data from the Health and Behaviors in Transition (HABIT) trial. Pregnant participants with pre-pregnancy body mass index [≥]25 kg/m2 wore a wrist-worn ActiGraph for seven consecutive days to estimate moderate-to-vigorous physical activity (MVPA), light physical activity (LPA), inactivity (IA), and sleep. We used compositional data analysis with an isometric log-ratio transformation to examine associations with depressive symptoms, quantified by the Edinburgh Postnatal Depression Scale (EPDS), adjusting for maternal age, socioeconomic status, and cohabitation status. G-computation was applied to estimate the marginal mean of depressive symptoms at specified durations of each behavior, with the remaining time reallocated proportionally among the other behaviors. Results: A total of 255 participants with median EPDS score of 5.0 (IQR, 2.0-8.0) had geometric mean 24-hour behaviors of MVPA = 75.5 min/day (5.2%), LPA = 311.2 min/day (21.6%), IA = 593.9 min/day (41.2%), and sleep = 459.4 min/day (31.9%). 24-hour time-use behaviors were not significantly associated with EPDS scores, and marginal mean EPDS scores showed minimal changes (MVPA 10 min: 6.51 (95% CI 4.22, 9.64) vs 40 min: 5.97 (95% CI 5.03, 6.97); sleep 300 min: 5.41 (95% CI 4.56, 6.22) vs 480 min: 5.81 (95% CI 5.23, 6.39)). Conclusions: In this relatively active pregnant population, 24-hour time-use behaviors were not associated with depressive symptoms. Future research can examine later trimesters or other populations that are more sedentary or more depressed.