title: Catalytic role of 2'-hydroxyl groups within a group II intron active site.
abstract: Domain 5 is an essential active-site component of group II intron ribozymes. The role of backbone substituents in D5 function was explored through synthesis of a series of derivatives containing deoxynucleotides at each position along the D5 strand. Kinetic screens revealed that eight 2'-hydroxyl groups were likely to be critical for activity of D5. Through two separate methods, including competitive inhibition and direct kinetic analysis, effects on binding and chemistry were distinguished. Depending on their function, important 2'-hydroxyl groups lie on opposite faces of the molecule, defining distinct loci for molecular recognition and catalysis by D5.
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Columbia University College of Physicians and Surgeons, New York, NY 10032, USA.
authors.forename: D L
authors.initials: DL
authors.lastname: Abramovitz
authors.forename: R A
authors.initials: RA
authors.lastname: Friedman
authors.forename: A M
authors.initials: AM
authors.lastname: Pyle
mesh_terms: D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D002384:Catalysis; D005091:Exons; D006860:Hydrogen Bonding; D017665:Hydroxyl Radical / Q000737:chemistry; D007438:Introns; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000737:chemistry / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D016337:RNA, Catalytic / Q000737:chemistry / Q000378:metabolism*
chemical_list: D009843:Oligoribonucleotides; D016337:RNA, Catalytic; D017665:Hydroxyl Radical; D012313:RNA