title: Identification of the RNA-binding sites of the triple gene block protein 1 of bamboo mosaic potexvirus.
abstract: The triple gene block protein 1 (TGBp1) encoded by open reading frame 2 of bamboo mosaic potexvirus (BaMV) was overexpressed in Escherichia coli and purified in order to test its RNA-binding activity. UV crosslinking assays revealed that the RNA-binding activity was present mainly in the soluble fraction of the refolded TGBp1. The binding activity was nonspecific and salt concentration-dependent: activity was present at 0-50 mM NaCl but was almost abolished at 200 mM. The RNA-binding domain was located by deletion mutagenesis to the N-terminal 3-24 amino acids of TGBp1. Sequence alignment analysis of the N-terminal 25 amino acids of the TGBp1 homologues of potexviruses identified three arginine residues. Arg-to-Ala substitution at any one of the three arginines eliminated most of the RNA-binding activity, indicating that they were all critical to the RNA-binding activity of the TGBp1 of BaMV.
authors.forename: C H
authors.initials: CH
authors.lastname: Wung
authors.forename: Y H
authors.initials: YH
authors.lastname: Hsu
authors.forename: D Y
authors.initials: DY
authors.lastname: Liou
authors.forename: W C
authors.initials: WC
authors.lastname: Huang
authors.forename: N S
authors.initials: NS
authors.lastname: Lin
authors.forename: B Y
authors.initials: BY
authors.lastname: Chang
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010942:Plant Viruses / Q000235:genetics / Q000378:metabolism*; D017863:Potexvirus / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D012313:RNA / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment; D017384:Sequence Deletion; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D016601:RNA-Binding Proteins; D014764:Viral Proteins; D012313:RNA