title: Arginine-rich motifs are not required for hepatitis delta virus RNA binding activity of the hepatitis delta antigen.
abstract: Hepatitis delta virus (HDV) replication and packaging require interactions between the unbranched rodlike structure of HDV RNA and hepatitis delta antigen (HDAg), a basic, disordered, oligomeric protein. The tendency of the protein to bind nonspecifically to nucleic acids has impeded analysis of HDV RNA protein complexes and conclusive determination of the regions of HDAg involved in RNA binding. The most widely cited model suggests that RNA binding involves two proposed arginine-rich motifs (ARMs I and II) in the middle of HDAg. However, other studies have questioned the roles of the ARMs. Here, binding activity was analyzed in vitro using HDAg-160, a C-terminal truncation that binds with high affinity and specificity to HDV RNA segments in vitro. Mutation of the core arginines of ARM I or ARM II in HDAg-160 did not diminish binding to HDV unbranched rodlike RNA. These same mutations did not abolish the ability of full-length HDAg to inhibit HDV RNA editing in cells, an activity that involves RNA binding. Moreover, only the N-terminal region of the protein, which does not contain the ARMs, was cross-linked to a bound HDV RNA segment in vitro. These results indicate that the amino-terminal region of HDAg is in close contact with the RNA and that the proposed ARMs are not required for binding HDV RNA. Binding was not reduced by mutation of additional clusters of basic amino acids. This result is consistent with an RNA-protein complex that is formed via numerous contacts between the RNA and each HDAg monomer.
authors.affiliation: Department of Microbiology and Immunology, Georgetown University Medical Center, Washington, DC, USA.
authors.forename: Leighton H
authors.initials: LH
authors.lastname: Daigh
authors.forename: Brittany L
authors.initials: BL
authors.lastname: Griffin
authors.forename: Ali
authors.initials: A
authors.lastname: Soroush
authors.forename: Murad R
authors.initials: MR
authors.lastname: Mamedov
authors.forename: John L
authors.initials: JL
authors.lastname: Casey
mesh_terms: D020816:Amino Acid Motifs; D001120:Arginine / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D004252:DNA Mutational Analysis; D003698:Hepatitis Delta Virus / Q000235:genetics / Q000502:physiology*; D035923:Hepatitis delta Antigens / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D012367:RNA, Viral / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion
chemical_list: D035923:Hepatitis delta Antigens; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D001120:Arginine