title: Hepatitis delta virus antigen is methylated at arginine residues, and methylation regulates subcellular localization and RNA replication.
abstract: Hepatitis delta virus (HDV) contains a circular RNA which encodes a single protein, hepatitis delta antigen (HDAg). HDAg exists in two forms, a small form (S-HDAg) and a large form (L-HDAg). S-HDAg can transactivate HDV RNA replication. Recent studies have shown that posttranslational modifications, such as phosphorylation and acetylation, of S-HDAg can modulate HDV RNA replication. Here we show that S-HDAg can be methylated by protein arginine methyltransferase (PRMT1) in vitro and in vivo. The major methylation site is at arginine-13 (R13), which is in the RGGR motif of an RNA-binding domain. The methylation of S-HDAg is essential for HDV RNA replication, especially for replication of the antigenomic RNA strand to form the genomic RNA strand. An R13A mutation in S-HDAg inhibited HDV RNA replication. The presence of a methylation inhibitor, S-adenosyl-homocysteine, also inhibited HDV RNA replication. We further found that the methylation of S-HDAg affected its subcellular localization. Methylation-defective HDAg lost the ability to form a speckled structure in the nucleus and also permeated into the cytoplasm. These results thus revealed a novel posttranslational modification of HDAg and indicated its importance for HDV RNA replication. This and other results further showed that, unlike replication of the HDV genomic RNA strand, replication of the antigenomic RNA strand requires multiple types of posttranslational modification, including the phosphorylation and methylation of HDAg.
authors.affiliation: Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, 2011 Zonal Ave., Los Angeles, CA 90033-1054, USA.
authors.forename: Yi-Jia
authors.initials: YJ
authors.lastname: Li
authors.forename: Michael R
authors.initials: MR
authors.lastname: Stallcup
authors.forename: Michael M C
authors.initials: MM
authors.lastname: Lai
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine / Q000378:metabolism; D045744:Cell Line, Tumor; D003698:Hepatitis Delta Virus / Q000502:physiology*; D035923:Hepatitis delta Antigens / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D008745:Methylation; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D012367:RNA, Viral / Q000096:biosynthesis*; D014162:Transfection; D014779:Virus Replication
chemical_list: D035923:Hepatitis delta Antigens; D012367:RNA, Viral; D001120:Arginine