Roles of the three major phosphorylation sites of hepatitis B virus core protein in viral replication.

Hepatitis B virus (HBV) core protein is a phosphoprotein. Its three major phosphorylation sites have been identified at the serine residues located at amino acids 157, 164, and 172. In order to investigate the role of core protein phosphorylation in HBV replication, these three serine residues were mutated to alanine to mimic nonphosphorylated serine or to glutamic acid to mimic phosphoserine. The nonphosphorylated core protein analog did not package the HBV pregenomic RNA, and the phosphorylated analog packaged the pregenomic RNA but failed to support viral DNA replication. These results indicate that the core protein phosphorylation may be important for pregenomic RNA packaging and that its dephosphorylation may be important for viral DNA replication. The individual roles of these three major phosphorylation sites in HBV replication were further investigated by being mutated to alanine in different combinations. The results showed that the serine residue at amino acid 157 was not essential for pregenomic RNA packaging, whereas the serine residues at amino acids 164 and 172 were more important. Furthermore, the serine residue at amino acid 157 was not essential for viral DNA replication or viral maturation.