The structure of Au clusters on the TiO2(110) surface and the cluster-substrate interaction were investigated by polarization-dependent total reflection fluorescence X-ray absorption fine structure (PTRF-XAFS). The TiO2(110) (1 脳 1) and (1 脳 2) surfaces were compared in terms of substrate structural effects on Au cluster structure. We could not find evidence for the direct Au-substrate bonding and observed that the substrate morphology did not significantly affect the size and structure of Au clusters. Finally, we precisely determined that the Au clusters favored a geometry with icosahedral Au55 symmetry. We discuss the possible interaction mechanism between Au and TiO2(110) that leads to these results.