NK cells can recognize target cells such as virus-infected and tumor cells through integration of activation and inhibitory receptors. Recognition by NK cells can lead to direct lysis of the target cell and production of the signature cytokine IFN-g. However, it is unclear whether stimulation through activation receptors alone is sufficient for IFN-g production. In this study, we show that NK activation receptor engagement requires additional signals for optimal IFN-g production, which could be provided by IFN-b or IL-12. Stimulation of murine NK cells with soluble Abs directed against NK1.1, Ly49H, Ly49D, or NKp46 required additional stimulation with cytokines, indicating that a range of activation receptors with distinct adaptor molecules require additional stimulation for IFN-g production. The requirement for multiple signals extends to stimulation with primary m157-transgenic target cells, which triggers the activation receptor Ly49H, suggesting that NK cells do require multiple signals for IFN-g production in the context of target cell recognition. Using quantitative PCR and RNA flow cytometry, we found that cytokines, not activating ligands, act on NK cells to express Ifng transcripts. Ly49H engagement is required for IFN-g translational initiation. Results using inhibitors suggest that the proteasome–ubiquitin–IKK–TPL2–MNK1 axis was required during activation receptor engagement. Thus, this study indicates that activation receptor–dependent IFN-g production is regulated on the transcriptional and translational levels.