title: Crystallization and X-ray diffraction analysis of the Trp/amber editing site of hepatitis delta virus (+)RNA: a case of rational design.
abstract: RNA editing by mammalian ADAR1 (Adenosine Deaminase Acting on RNA) is required for the life cycle of the hepatitis delta virus (HDV). Editing extends the single viral open reading frame to yield two protein products of alternate length. ADARs are believed to recognize double-stranded RNA substrates via a ;structure-based' readout mechanism. Crystals of 10-mer duplexes representing the HDV RNA-editing site diffracted to 1.35 A resolution, but suffered from merohedral twinning and averaging of the base registry. Expansion of the construct to include two flanking 3 x 1 internal loops yielded crystals in the primitive tetragonal space group P4(1)2(1)2 or P4(3)2(1)2. X-ray diffraction data were collected to 2.8 A resolution, revealing a unit cell with parameters a = 62.5, c = 63.5 A. The crystallization and X-ray analysis of multiple forms of the HDV RNA-editing substrate, encounters with common RNA crystal-growth defects and a strategy to overcome these problems are reported.
authors.affiliation: Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA.
authors.forename: Celeste
authors.initials: C
authors.lastname: MacElrevey
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Wedekind
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D003062:Codon; D003460:Crystallization; D003698:Hepatitis Delta Virus / Q000235:genetics*; D035923:Hepatitis delta Antigens / Q000235:genetics; D008969:Molecular Sequence Data; D017393:RNA Editing; D012367:RNA, Viral; D014961:X-Ray Diffraction
chemical_list: D003062:Codon; D035923:Hepatitis delta Antigens; D012367:RNA, Viral