33422-70-7Relevant articles and documents
Stability studies on the newly discovered cyclic form of tRNA N 6-threonylcarbamoyladenosine (ct6A)
Matuszewski, Michal,Sochacka, Elzbieta
, p. 2703 - 2706 (2014)
A cyclic form of N6-threonylcarbamoyladenosine bearing an oxazolone moiety (ct6A) was discovered very recently at the position 37 in several tRNA sequences. Our study on the synthesized 5′,3′, 2′-O-acetylated derivative of ct6A confirmed high stability of the modified nucleoside under physiological conditions (PBS buffer, pH 7.4) and revealed remarkable stability of the oxazolone ring in acidic (100 mM HCl, pH 1) and basic (0.1 mM NaOH, pH 10) conditions. This feature may allow for the post-synthetic conversion of t6A into ct6A in assembled oligoribonucleotides.
Synthesis and characterization of the native anticodon domain of E. coli tRNA(Lys): Simultaneous incorporation of modified nucleosides mnm5s2U, t6A, and pseudouridine using phosphoramidite chemistry
Sundaram, Mallikarjun,Crain, Pamela F.,Davis, Darrell R.
, p. 5609 - 5614 (2007/10/03)
The anticodon domain of E. coli tRNA(Lys) contains the hypermodified nucleosides mnm5s2U and t6A at positions 34 and 37, respectively, along with a more common ψ at position 39. The combination of these three nucleotides represents one of the most extensively modified RNA domains in nature. 2-Cyanoethyl diisopropylphosphoramidites of the hypermodified nucleosides mnm5s2U and t6A were each synthesized with protecting groups suitable for automated RNA oligonucleotide synthesis. The 17 nucleotide anticodon stem-loop of E. coli tRNA(Lys) was then assembled from these synthons using phosphoramidite coupling chemistry. Coupling efficiencies for the two hypermodified nucleosides and for pseudouridine phosphoramidite were all greater than 98%. A mild deprotection scheme was developed to accommodate the highly functionalized RNA. High coupling yields, mild deprotection, and efficient HPLC purification allowed us to obtain 1.8 mg of purified RNA from a 1 μmol scale RNA synthesis. Our efficient synthetic protocol will allow for biophysical investigation of this rather unique tRNA species wherein nucleoside modification has been shown to play a role in codon-anticodon recognition, tRNA aminoacyl synthetase recognition, and programmed ribosomal frameshifting. The human analogue, tRNA(Lys,3), is the specific tRNA primer for HIV-1 reverse transcriptase and has a similar modification pattern.