title: Synthesis of 2'-N-methylamino-2'-deoxyguanosine and 2'-N,N-dimethylamino-2'-deoxyguanosine and their incorporation into RNA by phosphoramidite chemistry.
abstract: The 2'-hydroxyl groups within RNA contribute in essential ways to RNA structure and function. Previously, we designed an atomic mutation cycle (AMC) that uses ribonucleoside analogues bearing different C-2'-substituents, including -OCH(3), -NH(2), -NHMe, and -NMe(2), to identify hydroxyl groups within RNA that donate functionally significant hydrogen bonds. To enable AMC analysis of the nucleophilic guanosine cofactor in the Tetrahymena ribozyme reaction and at other guanosines whose 2'-hydroxyl groups impart critical functional contributions, we describe here the syntheses of 2'-methylamino-2'-deoxyguanosine (G(NHMe)) and 2'-N,N-dimethylamino-2'-deoxyguanosine (G(NMe(2))) and their corresponding phosphoramidites. The key step in obtaining the nucleosides involved S(N)2 displacement of 2'-β-triflate from an appropriate guanosine derivative by methylamine or dimethylamine. We readily obtained the G(NMe(2)) phosphoramidite and incorporated it into RNA. However, the G(NHMe) phosphoramidite posed a significantly greater challenge due to lack of a suitable -2'-NHMe protecting group. After testing several strategies, we established that allyloxycarbonyl (Alloc) provided suitable protection for 2'-N-methylamino group during the phosphoramidite synthesis and the subsequent RNA synthesis. This work enables AMC analysis of guanosine's 2'-hydroxyl group within RNA.
authors.affiliation: Department of Biochemistry & Molecular Biology, The University of Chicago, 929 East 57th Street, MC 1028, Chicago, Illinois 60637, United States.
authors.forename: Qing
authors.initials: Q
authors.lastname: Dai
authors.forename: Raghuvir
authors.initials: R
authors.lastname: Sengupta
authors.forename: Shirshendu K
authors.initials: SK
authors.lastname: Deb
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Piccirilli
mesh_terms: D001483:Base Sequence; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D006860:Hydrogen Bonding; D015394:Molecular Structure; D009943:Organophosphorus Compounds / Q000737:chemistry*; D012313:RNA / Q000737:chemistry*; D012263:Ribonucleosides / Q000737:chemistry*
chemical_list: C570466:2'-N,N-dimethylamino-2'-deoxyguanosine; C570467:2'-N-methylamino-2'-deoxyguanosine; D009943:Organophosphorus Compounds; D012263:Ribonucleosides; C434331:phosphoramidite; D012313:RNA; D003849:Deoxyguanosine