Characterization of G-Quadruplex/Hairpin Transitions of RNAs by 19F NMR Spectroscopy

被引:20
|
作者
Granqvist, Lotta [1 ]
Virta, Pasi [1 ]
机构
[1] Univ Turku, Dept Chem, Turku 20014, Finland
基金
芬兰科学院;
关键词
bistable RNAs; fluorinated ligands; F-19 NMR spectroscopy; G-quadruplex RNAs; RNA structures; SECONDARY STRUCTURE REARRANGEMENTS; NUCLEIC-ACIDS; REDOR NMR; PROBES; SWITCHES; SENSOR; OLIGODEOXYNUCLEOTIDES; OLIGORIBONUCLEOTIDES; EQUILIBRIA; NUCLEOSIDE;
D O I
10.1002/chem.201602898
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
2'-O-[(4-Trifluoromethyl-triazol-1-yl)methyl] reporter groups have been incorporated into guanosine-rich RNA models (including a known bistable Qd/Hp RNA and two Grich regions of mRNA of human prion protein, PrP) and applied for the F-19 NMR spectroscopic characterization of plausible G-quadruplex/hairpin (Qd/Hp) transitions in these RNA structures. For the synthesis of the CF3-labeled RNAs, phosphoramidite building blocks of 2'-O-[(4-CF3-triazol-1-yl)methyl] nucleosides (cytidine, adenosine, and guanosine) were prepared and used as an integral part of the standard solid-phase RNA synthesis. The obtained F-19 NMR spectra supported the usual characterization data (obtained by UV-and CD-melting profiles and by H-1 NMR spectra of the imino regions) and additionally gave more detailed information on the Qd/Hp transitions. The molar fractions of the secondary structural species (Qd, Hp) upon thermal denaturation and under varying ionic conditions could be determined from the intensities and shifts of the F-19 NMR signals. For a well-behaved Qd/Hp transition, thermodynamic parameters could be extracted.
引用
收藏
页码:15360 / 15372
页数:13
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