This study analyzed the effect of rear-wheel position on seating comfort and mobility efficiency. Twelve randomly selected paraplegic wheelchair users participated in the study. Wheelchairs were tested in two rear-wheel positions while the users operated the wheelchair on a treadmill and while they worked on a computer. Propulsion efficiency, seating comfort, and propulsion qualities were registered at different loads during the treadmill session. During the computer session, pelvic position, estimated seating comfort, and estimated activity performance were measured. The change in rear-wheel position affected wheelchair ergonomics with respect to weight distribution (p < 0.0001) and seat inclination angle (position I = 5 and position II = 12). These changes had a significant effect on push frequency (p < 0.05) and stroke angle (p < 0.05) during wheelchair propulsion. We found no consistent effect on mechanical efficiency, estimated exertion, breathlessness, seating comfort, estimated propulsion qualities, pelvic position, or activity performance.