The Small Delta Antigen of Hepatitis Delta Virus Is an Acetylated Protein and Acetylation of Lysine 72 May Influence Its Cellular Localization and Viral Rna Synthesis

Abstract

Hepatitis delta virus (HDV) is a single-stranded RNA virus that encodes two viral nucleocapsid proteins named small and large form hepatitis delta antigen (S-HDAg and L-HDAg). The S-HDAg is essential for viral RNA replication while the L- HDAg is required for viral assembly. In this study, we demonstrated that HDAg are acetylated proteins. Metabolic labeling with [H-3]acetate revealed that both forms of HDAg could be acetylated in vivo. The histone acetyltransferase ( HAT) domain of cellular acetyltransferase p300 could acetylate the full-length and the N- terminal 88 amino acids of S-HDAg in vitro. By mass spectrometric analysis of the modified protein, Lys-72 of S-HDAg was identified as one of the acetylation sites. Substitution of Lys-72 to Arg caused the mutant S-HDAg to redistribute from the nucleus to the cytoplasm. The mutant reduced viral RNA accumulation and resulted in the earlier appearance of L-HDAg. These results demonstrated that HDAg is an acetylated protein and mutation of HDAg at Lys-72 modulates HDAg subcellular localization and may participate in viral RNA nucleocytoplasmic shuttling and replication. (C) 2003 Elsevier Inc. All rights reserved