Participation of 9 to 11-year-old children in physical education lessons (PEL) contributes to a significantly higher duration of moderate-to-vigorous physical activity (MVPA) during the school day and, in overweight/obese girls and normal-weight boys, to an increase in overall daily MVPA as shown by previous research. However, it is not known whether this increase in MVPA is at the expense of light physical activity (LPA) or sedentary behaviour (SED). SED, LPA, and MVPA were assessed in 338 schoolchildren aged 9-11 years (50.3% girls; 29.6% overweight/obese) over two school days (with and without a PEL) using a triaxial accelerometer during various segments of the school day. SED, LPA, and MVPA were quantified based on the duration of the activity (minutes). Participation in PEL led to significantly higher school MVPA in the overweight/obese and normal-weight girls and boys (p < 0.005) compared to MVPA of those children on the school day without PEL. Participation in PEL led to a significantly higher overall daily MVPA duration compared to that during the day without PEL for the overweight/obese girls (p < 0.05), normal-weight girls (p < 0.05) and boys (p < 0.005). Participation in PEL contributed not only to significantly higher LPA in the normal-weight girls and boys (p < 0.01) during the school day but also reduced school-time SED in the overweight/obese children (p < 0.01) and normal-weight girls (p < 0.005). Moreover, participation in PEL significantly reduced the overall daily SED in the normal-weight children and overweight/obese boys (p < 0.05). Adding one PEL to the daily school routine appears to be a promising strategy for effectively reducing SED in children.