This paper presents the effects of bearing locations on the mechanical characteristics of a multi-stepped spindle system related to bearing fatigue life, natural frequency, and static stiffness. The multi-stepped spindle is supported by a pair of tapered roller bearings (TRBs) and subjected to radial loading. To solve the equilibrium equation of the spindle system which is inherently statically-indeterminate, this study adopts an integrated shaft-bearing model, where the spindle is modelled by the finite shaft elements and the supporting TRBs are modelled by the five degrees-of-freedom TRB model developed by the authors. An iterative computational method is used to estimate the spindle deflection coupled with bearing deflections, and afterwards the bearing stiffness and internal contact loads of rolling elements are computed. The bearing fatigue life based on the ISO standard and the first natural frequency of the spindle system are evaluated with the spindle-bearing model. The influences of bearing locations on the static stiffness and natural frequency of the spindle, and the fatigue life of TRBs are rigorously investigated. The numerical results show the noticeable effects of bearing locations on the spindle system characteristics. The presented results provide a comprehensive assessment to aid for design optimization of spindle-TRB system.