title: The Escherichia coli tRNA-guanine transglycosylase can recognize and modify DNA.
abstract: tRNA-guanine transglycosylase (TGT) catalyzes the exchange of queuine (or a precursor) for guanine 34 in tRNA. The minimal RNA recognition motif for TGT has been found to involve a UGU sequence in the anticodon loop of the queuine-cognate tRNAs. Recent studies have shown that the enzyme is capable of recognizing the UGU sequence in alternative contexts (Kung, F. L., Nonekowski, S., and Garcia, G. A. (2000) RNA 6, 233-244) and have investigated the role of the first U of the UGU sequence in tRNA recognition by TGT (Nonekowski, S. T., and Garcia, G. A. (2001) RNA 7, 1432-1441). The TGT reaction involves the breakage and re-formation of a glycosidic bond. To rule out a potential chemical mechanism involving the 2'-hydroxyl at position 34, we synthesized and evaluated an RNA minihelix with 2'-deoxy-G at 34. The high level of activity exhibited by this analogue indicates that the 2'-hydroxyl of G(34) is not required for catalysis. Furthermore, we find that TGT can recognize analogues composed entirely of DNA, but only when 2'-deoxyuridines replace the thymidines in the DNA. The requirement for uridine bases for recognition is perhaps not surprising given the UGU recognition motif for TGT. However, it is not clear if the uracil requirement is due to specific recognition by TGT or due to the effect of uracils on the conformation of the oligonucleotide.
authors.affiliation: Interdepartmental Program in Medicinal Chemistry, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109-1065, USA.
authors.forename: Susanne T
authors.initials: ST
authors.lastname: Nonekowski
authors.forename: Fan-Lu
authors.initials: FL
authors.lastname: Kung
authors.forename: George A
authors.initials: GA
authors.lastname: Garcia
mesh_terms: D001483:Base Sequence; D002384:Catalysis; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D003857:Deoxyuridine / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry; D010430:Pentosyltransferases / Q000378:metabolism*; D011485:Protein Binding; D012313:RNA / Q000378:metabolism; D013936:Thymidine / Q000737:chemistry; D014498:Uracil / Q000737:chemistry
chemical_list: D009841:Oligonucleotides; D014498:Uracil; D012313:RNA; D004247:DNA; D010430:Pentosyltransferases; C020837:queuine tRNA-ribosyltransferase; D013936:Thymidine; D003857:Deoxyuridine