Participation in youth sport is positively associated with physical fitness and performance. The purpose of the current study was to examine if physical fitness measures during childhood and early adolescence predicted high school sport participation. Participants included youth in the Michigan State University Motor Performance Study. Measures consisted of the endurance shuttle run, Wells sit-and-reach, jump and reach, standing long jump, agility shuttle run and 30-yard dash. Individual, generalized estimating equation models were used to estimate the association between each measure and the odds of participating in high school sport. For both sexes, the endurance shuttle run and agility shuttle run were related to sport participation. For boys only, the long jump and 30-yard dash were also associated with sport participation. Better physical fitness and performance across several dimensions in childhood, including those related to motor skills, enhanced the likelihood of sport participation in high school.