The hepatitis B virus X-gene product trans-activates both RNA polymerase II and III promoters

Abstract

The transcriptional regulatory activity of the human hepatitis B virus (HBV) X-gene product was investigated. We demonstrate a new property for the HBV X-gene, the strong transcriptional trans-activation of promoters for class III genes. The stimulation of RNA polymerase III (pol III) as well as pol II promoters is shown in cells transiently transfected with the X-gene, and after its stable integration into hepatocytes. We demonstrate that X-gene containing cells stimulate the frequency of pol III transcription initiation by 20- to 40-fold, and accelerate the rate of formation of stable pol III initiation complexes in a manner indistinguishable from that of adenovirus E1a protein. Since the transcription factor TFIIIC has been shown to be limiting in the formation of stable pol III initiation complexes, template commitment experiments were performed which titrate the level of this factor in extracts. We show that X-protein containing extracts are far more efficient in forming stable pol III preinitiation complexes that cannot be competed away upon addition of a second template, indicating that TFIIIC is very probably a target of the X-protein. Thus, the HBV X-protein is apparently a member of a family of trans-activators capable of stimulating both pol II and III promoters, which includes the adenovirus E1a-protein and SV40 t antigen.