Hepatitis delta virus genome replication: a polyadenylated mRNA for delta antigen

Hepatitis delta virus (HDV) replicates its genome in the nucleus of an infected cell. However, an unsolved problem has been the identification in the cytoplasm of a putative mRNA for the synthesis of the only virus-coded protein, the delta antigen. We now report the characterization of an 800-base RNA that is cytoplasmic, polyadenylated, and antigenomic and that should direct the translation of the delta antigen. This RNA was about 500 times less abundant than full-length genomic RNA. We mapped the predominant 5' terminus and also the 3' site at which the poly(A) is added. At a point 15 to 20 bases upstream of the poly(A) addition site is the sequence AAUAAA, which could have been used as a signal for the polyadenylation. When an infectious cDNA clone of the whole HDV genome was changed at this site to UUUAAA, the clone was no longer infectious and it was unable to direct the synthesis of the delta antigen. These findings provided additional evidence that the polyadenylated RNA was at least the predominant method for the expression of the delta antigen. Apparently the HDV RNA was processed as if it were a host mRNA polymerase II transcript, although this did not necessarily indicate that HDV RNA was transcribed with this enzyme.