The main aim of the present paper is to assess the fatigue lifetime of ductile cast iron knuckles as one of the critical components of an automotive steering system. To this end, a real driving path, including various maneuvers, such as acceleration, braking, cornering, and moving on various types of road roughness, was considered. Different load histories, which are applied on various joints of the component (i.e., lower control arm, steering linkage, and Macpherson strut), were extracted through Multi-Body Dynamics (MBD) analysis of a full vehicle model. The achievements of previous studies have proved that the steering knuckle fails from the steering linkage and due to the rotational motion. Therefore, only this destructive load history was considered in future analyses of the present study. The CAD model was created using Coordinate Measuring Machine (CMM) data and some corrections in CATIA software. Furthermore, transient dynamic analysis was performed, and the time history of von Misses equivalent stress was obtained at the root of the steering linkage (which is exactly the location of failure based on the laboratory data as well as finite element simulations validated by the author in the previous studies). To predict the fatigue life of a component, two different methodologies were considered. Firstly, some well-known criteria were employed for equalization of load spectrum to a Constant Amplitude Loading (CAL). Then, fatigue analysis under sinusoidal loading was performed. Secondly, the fatigue life of the component considering Variable Amplitude Loading (VAL) was predicted using the Critical Plane Method (CPM), employing the Rain-flow cycle counting technique, and utilizing the Miner–Palmgren damage accumulation rule. Eventually, to evaluate the prediction accuracy of different methodologies, the obtained results were compared with the full-scale axial variable amplitude fatigue test which was performed by the corresponding author. The results indicated that the prediction of variable amplitude fatigue lifetime by Finite Element (FE) analysis in the time domain has about 21% error compared to reality. Additionally, the relative error between the results obtained from two different methodologies is about 20%, which is acceptable due to the scattering of the fatigue phenomenon results, the complex geometry of the part, and the complicated loading.