title: Synthesis and incorporation of the phosphoramidite derivative of 2'-O-photocaged 3'-s-thioguanosine into oligoribonucleotides: substrate for probing the mechanism of RNA catalysis.
abstract: Oligoribonucleotides containing 3'-S-phosphorothiolate linkages possess properties that can reveal deep mechanistic insights into ribozyme-catalyzed reactions. "Photocaged" 3'-S- RNAs could provide a strategy to stall reactions at the chemical stage and release them after assembly steps have occurred. Toward this end, we describe here an approach for the synthesis of 2'-O-(o-nitrobenzyl)-3'-thioguanosine phosphoramidite starting from N(2)-isobutyrylguanosine in nine steps with 10.2% overall yield. Oligonucleotides containing the 2'-O-(o-nitrobenzyl)-3'-S-guanosine nucleotide were then constructed, characterized, and used in a nuclear pre-mRNA splicing reaction.
authors.affiliation: Department of Biochemistry & Molecular Biology, ‡Department of Chemistry, and §Department of Molecular Genetics & Cell Biology, University of Chicago , Chicago, Illinois 60637, United States.
authors.forename: Nan-Sheng
authors.initials: NS
authors.lastname: Li
authors.forename: Nicole
authors.initials: N
authors.lastname: Tuttle
authors.forename: Jonathan P
authors.initials: JP
authors.lastname: Staley
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Piccirilli
mesh_terms: D006151:Guanosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D015335:Molecular Probes / Q000138:chemical synthesis* / Q000737:chemistry; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000737:chemistry*; D009943:Organophosphorus Compounds / Q000138:chemical synthesis*; D010710:Phosphates / Q000138:chemical synthesis* / Q000737:chemistry; D012326:RNA Splicing; D016337:RNA, Catalytic / Q000737:chemistry*
chemical_list: C000592676:2'-O-(o-nitrobenzyl)-3'-thioguanosine phosphoramidite; C465503:3'-S-phosphorothiolate; D015335:Molecular Probes; D009843:Oligoribonucleotides; D009943:Organophosphorus Compounds; D010710:Phosphates; D016337:RNA, Catalytic; C434331:phosphoramidite; D006151:Guanosine