title: S-Geranyl-2-thiouridine wobble nucleosides of bacterial tRNAs; chemical and enzymatic synthesis of S-geranylated-RNAs and their physicochemical characterization.
abstract: Recently, highly lipophilic S-geranylated derivatives of 5-methylaminomethyl-2-thiouridine (mnm5geS2U) and 5-carboxymethylaminomethyl-2-thiouridine (cmnm5geS2U) were found at the first (wobble) anticodon position in bacterial tRNAs specific for Lys, Glu and Gln. The function and cellular biogenesis of these unique tRNAs remain poorly understood. Here, we present one direct and two post-synthetic chemical routes for preparing model geS2U-RNAs. Our experimental data demonstrate that geS2U-RNAs are more lipophilic than their parent S2U-RNAs as well as non-modified U-RNAs. Thermodynamic studies revealed that the S-geranyl-2-thiouridine-containing RNA has higher affinity toward complementary RNA strand with G opposite the modified unit than with A. Recombinant tRNA selenouridine synthase (SelU) exhibits sulfur-specific geranylation activity toward model S2U-RNA, which is composed of the anticodon-stem-loop (ASL) from the human tRNALys3sequence. In addition, the presence of magnesium ions is required to achieve appreciable geranylation efficiencies.
authors: Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Department of Bioorganic Chemistry, Sienkiewicza 112, 90-363 Lodz, Poland.
authors: Malgorzata
authors: M
authors: Sierant
authors: Institute of Organic Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego 116, 90-924 Lodz, Poland.
authors: Grazyna
authors: G
authors: Leszczynska
authors: Institute of Organic Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego 116, 90-924 Lodz, Poland.
authors: Klaudia
authors: K
authors: Sadowska
authors: Institute of Organic Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego 116, 90-924 Lodz, Poland.
authors: Agnieszka
authors: A
authors: Dziergowska
authors: Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Department of Bioorganic Chemistry, Sienkiewicza 112, 90-363 Lodz, Poland.
authors: Michal
authors: M
authors: Rozanski
authors: Institute of Organic Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego 116, 90-924 Lodz, Poland.
authors: Elzbieta
authors: E
authors: Sochacka
authors: Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Department of Bioorganic Chemistry, Sienkiewicza 112, 90-363 Lodz, Poland bnawrot@cbmm.lodz.pl.
authors: Barbara
authors: B
authors: Nawrot
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry; D020029:Base Pairing; D001483:Base Sequence; D001665:Binding Sites; D005960:Glucosides / Q000138:chemical synthesis; D008274:Magnesium / Q000737:chemistry; D012329:RNA, Bacterial / Q000138:chemical synthesis / Q000235:genetics*; D012343:RNA, Transfer / Q000138:chemical synthesis / Q000235:genetics*; D013816:Thermodynamics; D013891:Thiouridine / Q000031:analogs & derivatives* / Q000737:chemistry*; D044366:Transition Temperature
chemical_list: D005960:Glucosides; D012329:RNA, Bacterial; D013891:Thiouridine; D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases; D008274:Magnesium