title: Design, Synthesis, and Properties of Phosphoramidate 2',5'-Linked Branched RNA: Toward the Rational Design of Inhibitors of the RNA Lariat Debranching Enzyme.
abstract: Two RNA fragments linked by means of a 2',5' phosphodiester bridge (2' hydroxyl of one fragment connected to the 5' hydroxyl of the other) constitute a class of nucleic acids known as 2'-5' branched RNAs (bRNAs). In this report we show that bRNA analogues containing 2'-5' phosphoramidate linkages (bN-RNAs) inhibit the lariat debranching enzyme, a 2',5'-phosphodiesterase that has recently been implicated in neurodegenerative diseases associated with aging. bN-RNAs were efficiently generated using automated solid-phase synthesis and suitably protected branchpoint building blocks. Two orthogonally removable groups, namely the 4-monomethoxytrityl (MMTr) group and the fluorenylmethyl-oxycarbonyl (Fmoc) groups, were evaluated as protecting groups of the 2' amino functionality. The 2'-N-Fmoc methodology was found to successfully produce bN-RNAs on solid-phase oligonucleotide synthesis. The synthesized bN-RNAs resisted hydrolysis by the lariat debranching enzyme (Dbr1) and, in addition, were shown to attenuate the Dbr1-mediated hydrolysis of native bRNA.
authors.affiliation: Department of Chemistry, McGill University , 801 Sherbrooke Street West, Montréal, Québec H3A 0B8, Canada.|Department of Life Science, Tokyo Institute of Technology , 4259 Nagatsuta, Midoriku, Yokohama, Kanagawa 226-8501, Japan.
authors.forename: Nobuhiro
authors.initials: N
authors.lastname: Tago
authors.affiliation: Department of Chemistry, McGill University , 801 Sherbrooke Street West, Montréal, Québec H3A 0B8, Canada.
authors.forename: Adam
authors.initials: A
authors.lastname: Katolik
authors.affiliation: Department of Veterans Affairs, South Texas Veterans Health Care System , San Antonio, Texas 78229, United States.
authors.forename: Nathaniel E
authors.initials: NE
authors.lastname: Clark
authors.affiliation: Department of Veterans Affairs, South Texas Veterans Health Care System , San Antonio, Texas 78229, United States.|Departments of Biochemistry and Biomolecular Chemistry, University of Wisconsin-Madison , 433 Babcock Dr., Madison, Wisconsin 53706, United States.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Montemayor
authors.affiliation: Department of Life Science, Tokyo Institute of Technology , 4259 Nagatsuta, Midoriku, Yokohama, Kanagawa 226-8501, Japan.
authors.forename: Kohji
authors.initials: K
authors.lastname: Seio
authors.affiliation: Department of Life Science, Tokyo Institute of Technology , 4259 Nagatsuta, Midoriku, Yokohama, Kanagawa 226-8501, Japan.
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Sekine
authors.affiliation: Department of Veterans Affairs, South Texas Veterans Health Care System , San Antonio, Texas 78229, United States.|Department of Biochemistry, University of Texas Health Science Center , 7703 Floyd Curl Drive, San Antonio, Texas 78229, United States.
authors.forename: P John
authors.initials: PJ
authors.lastname: Hart
authors.affiliation: Department of Chemistry, McGill University , 801 Sherbrooke Street West, Montréal, Québec H3A 0B8, Canada.
authors.forename: Masad J
authors.initials: MJ
authors.lastname: Damha
mesh_terms: D000577:Amides / Q000737:chemistry*; D006801:Humans; D009690:Nucleic Acid Conformation; D010756:Phosphoric Acids / Q000737:chemistry*; D012313:RNA / Q000138:chemical synthesis* / Q000737:chemistry* / Q000378:metabolism; D012316:RNA Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D012326:RNA Splicing; D060327:Solid-Phase Synthesis Techniques
chemical_list: D000577:Amides; D010756:Phosphoric Acids; D012313:RNA; C011067:phosphoramidic acid; D012316:RNA Nucleotidyltransferases; C048135:lariat debranching enzyme