A transactivating function encoded in the hepatitis B virus X gene is conserved in the integrated state

Abstract

Recently we have shown that the hepatitis B virus (HBV) X gene encodes a transactivating factor. Here we report on the construction of a series of HBx expression plasmids which were tested for a transactivating function by cotransfections with a plasmid expressing the CAT gene under control of the SV40 early promoter. One of the plasmids, expressing a HBx specific protein, is shown to transiently transactivate CAT gene expression after cotransfection into the human cell line CC113. Furthermore, a cloned integrated HBV DNA sequence is also shown to transactivate several viral promoters and LTRs. By sequence analyses we can demonstrate that only the HBx ORF is intact and that it is fused to an ORF of at least 228 bp in the flanking cellular DNA. The HBx gene is cotranscribed with the flanking cellular DNA, resulting in a RNA of approximately 10 kb. By subcloning of the integrate we can demonstrate that the presence of the HBx gene and its expression by the HBV enhancer and/or the HBx promoter is required for the transactivating function of the integrated HBV DNA.