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The X protein of hepatitis B virus (HBV) has been shown to be a trans-activator for viral and cellular genes. Amino acid sequences in X protein were found to be highly homologous to functionally essential sequences in the "Kunitz domain," characteristic of Kunitz-type serine protease inhibitors. Mutations at these sequences completely abolished trans-activation. Consequently, HBV X protein resembles a serine protease inhibitor or its analogue, and may bring about trans-activation by activating certain transcriptional factors through proteolytic cleavage alteration.

To examine the transforming potential of the X gene product of hepatitis B virus (HBV), the X-gene-containing region (referred to as the HBx region) was introduced into mouse NIH3T3 cells. Each transformed cell line expressed X-coding mRNA at a different level. A positive correlation was found between the level of X-coding mRNA and the saturation density of the cells. The HBx-transformed cell lines exhibited X protein production and tumor formation in nude mice. The function of HBV in oncogenesis may involve the continuous expression of the X-gene-coded product in the HBV DNA-integrated cells.

We have employed a short-term transfection assay system in which we monitored the transient expression of the chloramphenicol acetyltransferase (CAT) gene linked to the promoter region of the normal and mutant T24 H-ras1 gene or the human epsilon-globin gene in Chinese hamster lung (CHL) cells or cells derived from them which carry and express one or the other of the polyoma virus early genes. Our findings can be summarized as follows: (i) The mutant T24 H-ras1 promoter region behaves as a stronger promoter than the H-ras1 gene in all these types of cells as well as in rat 208F fibroblast cells. (ii) In CHL cells expressing the polyoma large T antigen the normal and mutant T24 Ha-ras1 promoters are not trans-activated in these cells and only a 2.5-fold activation of the epsilon-globin promoter is observed. (iii) In cells expressing the polyoma middle T antigen both the normal and mutant H-ras1 are trans-activated whereas transcription from the epsilon-globin promoter is not affected when compared to the normal CHL cells. (iv) In cells expressing the polyoma small T antigen the normal and mutant H-ras1 as well as the epsilon-globin promoters are trans-activated. We suggest from these data that a tissue-specific element exists in the promoter region of the H-ras1 gene and that the polyoma middle and small T antigens trigger the expression of proteins that trans-activate these promoters.