The rear outside wheel of an over-the-road 18-wheel tractor-trailer failed at its bolt holes, permitting the tire and wheel to separate from the hub. Failure analysis was conducted using photographic examination of actual fracture surfaces and SEM examination of fracture replicas. The examinations indicated that fatigue cracks had originated at the rim backside and propagated to the rim midsection. Catastrophic failure occurred at a final overland. Fatigue fracture of the wheel rim was attributed to cyclic loading created by improper wheel mounting of a spare tire.