The envelope (Env) of Jaagsiekte sheep retrovirus (JSRV) functions as an oncoprotein. One of the mechanisms of JSRV-induced cell transformation that has been proposed for epithelial cells involves JSRV Env binding Hyaluronidase 2 (the JSRV receptor), thereby inducing its degradation and allowing the release and activation of RON tyrosine kinase which is normally suppressed by HYAL-2. In this study, we report that HYAL-2 and RON are not critical for the JSRV Env-induced transformation of the rat epithelial cell line IEC-18, while the cytoplasmic tail of the JSRV Env is critical to transform this cell line. We have also determined that RON can associate with the JSRV Env under normal and stringent conditions. In addition, the cytoplasmic tail of the JSRV and the enJS5F16 (non oncogenic JSRV-related endogenous retrovirus) Env proteins appears to have a major influence on the activation status of RON. Thus, it appears that the interaction of the JSRV Env with RON is more complex than previously thought and requires further investigation.