MVM(p) NS-2 protein expression is required with NS-1 for maximal cytotoxicity in human transformed cells

Abstract

The parvovirus-encoded nonstructural (NS) proteins have been implicated in the cytopathogenicity of these agents. Although protein NS-1 of minute virus of mice (MVM) has been shown to be toxic, little is known about the role of NS-2 in this process. In order to determine the contribution of NS-1 and NS-2 to cytotoxicity, we took advantage of an expression system controlled by the mouse mammary tumor virus promoter which responds to glucocorticoid stimulation and which controls the expression of both MVM(p) NS proteins. Different mutations were introduced in NS genes so as to affect the NS-1 or NS-2 protein. Neoplastic human cell lines expressing only NS-1 protein after induction by dexamethasone undergo a smaller lethality compared to lines expressing both wild-type proteins. Mutations that were introduced in NS-1 coding sequence and did not affect NS-2 were found to drastically suppress the cytotoxic effect. It is concluded that the NS-2 protein has little cytotoxic activity by itself but is required for the full expression of the viral cytopathic effect on transformed human cells. Furthermore these results lead us to suggest that the NS-2 cytotoxic domain is localized in the amino-terminal portion of the protein.