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The interactions between movement behaviors (physical activity, screen time, and sleep) affect the health of preschool children. Therefore, we examined the status of adherence to combinations of 24-hour movement guidelines (24-h MG) in Japanese preschool children and determined the associations between overweight/obesity and adherence to these 24-h MG. This cross-sectional study was conducted with 421 children aged 3–5 years (216 boys and 199 girls) living in the northeastern region of Japan. To evaluate the 24-h MG, physical activity over one week was measured using a three-axis accelerometer. For screen time and sleep duration, a questionnaire survey was conducted. Children who failed to meet all the 24-h MG had a higher probability of overweight/obesity than those who met all the 24-h MG (odds ratio 1.139, 95% confidence interval: 1.009, 1.285). The percentage of adherence to the 24-h MG was 91.6% for physical activity, 82.5% for sleep duration, and 33.7% for screen time, and only 21.5% of the children adhered to all three areas of the guidelines. Our findings have important implications for developing public health policies and effective intervention programs for preschool children.