Interactions among nutrients have been widely recognized in plants and play important roles in crop growth and yield formation. However, the interplay of Cu and N in rice plants is not yet clear. In this study, rice plants were grown with different combinations of Cu and N supply. The effects of Cu-N interaction on the growth, yield production, Cu and N transport, and gene expression levels were analyzed. The results showed that the effect of N supply on rice growth and yield formation was more pronounced than that of Cu supply. The Cu supply significantly improved the uptake of N (by 9.52–30.64%), while the N supply significantly promoted the root-to-shoot translocation of Cu (by 27.28–38.45%) and distributed more Cu (1.85–19.16%) into the shoots and leaves. The results of qRT-PCR showed that +Cu significantly up-regulated the expression levels of both NO3− and NH4+ transporter genes OsNRTs and OsAMTs, including OsNRT1.1B, OsNRT2.1, OsNRT2.3a, OsNRT2.4, OsAMT1.2, OsAMT1.3, and OsAMT3.1. Meanwhile, +N significantly up-regulated the expression levels of Cu transporter genes OsHMA5 and OsYSL16. In addition, the supply of Cu up-regulated the expression levels of OsGS1;2, OsGS2, and OsNADH-GOGAT to 12.61-, 6.48-, and 6.05-fold, respectively. In conclusion, our study demonstrates a synergistic effect between Cu and N in rice plants. It is expected that our results would be helpful to optimize the application of N and Cu fertilizers in agriculture.