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270 mhz proton nmr studies of four branched tetraribonucleotides which are formed as the lariat branch point in the pre messenger rna processing reactions splicing


, : 270 mhz proton nmr studies of four branched tetraribonucleotides which are formed as the lariat branch point in the pre messenger rna processing reactions splicing. Tetrahedron 44(2): 571-590

1H-NMR conformational studies of four title branched ribonucleotides, formed as a local structure in the splicing intron, have been carried out by assigning 1H resonances by 2D NMR such as triple Relay-COSY and 31P/1H correlation experiments and then observing the temperature-dependent (7-80.degree.C) (i) chemical shifts of aromatic and sugar protons, and (ii) shifts of N=S equilibrium of different sugar moieties. Detailed 2D NOE (NOESY) experiments have subsequently furnished informations regarding the spatial proximities of different protons in these molecules. These studies suggest a coherent conformational picture of the four title branched tetraribonucleotides: (a) the branch-point adenosine (A*) and its 3'-terminal pyrimidine residue are N-N stacked; (b) the A* is coplanar with its 2'-terminal guanosine residue; (c) the nucleotide (A or U) at the 5'-terminus of the A* is S-S stacked with the 2'-terminal guanosine residue and it is also S-N stacked to the A*; (d) the glycosidicbonds of the 3'-terminal pyrimidine residue(C or U) and 5'-terminal residue (A) are in anti and syn conformations respectively, whereas the A* and the 2'-terminal guanosine residues have approximately an equal population of syn and anti conformers. These conformationalfeatures of the branched tetraribonucleotides are reminischent of a distorted linear 3'-5' linked single stranded A-RNA helix fragment.

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