This paper investigates the failure mechanism of the rear drive shaft in a modified pickup truck which had operated for about 3000 km. The investigation included macroscopic and microscopic evaluation to document the morphologies of the fracture surface, measurement of the material composition, metallographic preparation and examination, mechanical testing, and finite element modelling and calculations. The results obtained suggest that rotation-bending fatigue was the primary cause of the drive shaft failure. The crack initiation is located in the root of the machined threads on the drive shaft surface and expanded along the side of the machining line surface. The main cause of fatigue cracks is attributable to a high stress concentration owing to a large unilateral bending impact under overload. Meanwhile, the bidirectional torsional force also produces a higher stress concentration and thus accelerates the fatigue crack to expand radially along the surface. Furthermore, the hardness of the central section of the drive shaft was marginally below standard. This deficiency results in harm to the bearings and other mechanical components, as well as expediting the enlargement of cracks. Finite element analysis revealed significant contact stress between the bearing and drive shaft, with stress levels exceeding the fatigue limit stress of the parent material. This highlights the need for reevaluation of the heat treatment process and vehicle loading quality to enhance the drive shaft’s longevity.