Some steps of the soil nitrogen (N) cycle are sensitive to environmental pressures like soil moisture or contamination, which are expected to evolve during the next decades. Individual stresses have been well studied, but their combination is not yet documented. In this work, we aimed at assessing the importance of the soil moisture on the impact of copper (Cu) contaminations on the N cycling soil function using the potential nitrification activities (PNA) as bioindicator. A two-step experiment was performed. First, a loamy soil was incubated 5 weeks in either 30, 60, or 90% of its water holding capacity (WHC) or alternating drought and rewetting periods. Thereafter, soil samples were exposed to a gradient of Cu concentrations through a bioassay involving nitrification. The dose-response curves of PNA in function of added Cu were modeled to calculate the effective Cu concentrations, namely ECx with x being the percentage of PNA inhibition. These values were then compared between experimental conditions to highlight differences in threshold values. The preincubation moisture treatments significantly affected the PNA responses to the secondary Cu stress with, for instance, hormetic responses in all cases except for the dry-rewetting treatment. Small PNA inhibitions were estimated for high Cu doses in the soils with low water contents (30% WHC) or submitted to dry-rewetting cycles, contrarily to the patterns observed for the soils with high water contents (90% WHC) or submitted to a single period of drought. Overall, significant differences were found in estimated ECx values between moisture treatments.