Past major earthquakes have witnessed extensive pile-supported transmission tower-line systems (PSTLs) failing, which highlights the great susceptibility of PSTLs to damage from earthquakes. However, a comprehensive literature review in seismic analysis of PSTLs reveals that two critical aspects, i.e. soil–structure interaction (SSI) and depth-varying spatial ground motions (DVSGMs), have been completely overlooked in numerous previous studies. To be specific, pile-supported transmission towers are generally assumed to be fixed to the ground surface and are excited using uniform ground motion inputs. Such analytical schemes may result in severe misestimates of seismic response predictions of PSTLs. Within this context, the main objective of the present study is to accurately assess the seismic performance of PSTLs by considering SSI and using DVSGMs as inputs. For this purpose, an existing prototype PSTL is firstly selected and the corresponding three-dimensional finite element model is created in ABAQUS software, in which SSI is simulated by the Beam on Nonlinear Winkler Foundation (BNWF) model. Then, the three-dimensional DVSGMs are stochastically synthesized based on the computed ground motion transfer functions (GMTFs) of local sites. Next, seismic performance of the PSTL with SSI, including dynamic responses, ultimate bearing capacity, and failure mechanism, is assessed using the generated DVSGMs as seismic inputs. Finally, a parametric study is conducted to comprehensively examine and discuss the influences of SSI, seismic excitation types, coherence loss, and local site conditions on seismic performance of the PSTL. Numerical results show that seismic performance of PSTLs can be affected significantly by the above mentioned influencing factors. This research is expected to offer a meaningful reference to seismic performance assessments of PSTLs.