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The essential role of stacking adenines in a two-base-pair RNA kissing complex.

Authors
  • Stephenson, William
  • Asare-Okai, Papa Nii
  • Chen, Alan A
  • Keller, Sean
  • Santiago, Rachel
  • Tenenbaum, Scott A
  • Garcia, Angel E
  • Fabris, Daniele
  • Li, Pan T X
Type
Published Article
Journal
Journal of the American Chemical Society
Publisher
American Chemical Society
Publication Date
Apr 17, 2013
Volume
135
Issue
15
Pages
5602–5611
Identifiers
DOI: 10.1021/ja310820h
PMID: 23517345
Source
Medline
License
Unknown

Abstract

In minimal RNA kissing complexes formed between hairpins with cognate GACG tetraloops, the two tertiary GC pairs are likely stabilized by the stacking of 5'-unpaired adenines at each end of the short helix. To test this hypothesis, we mutated the flanking adenines to various nucleosides and examined their effects on the kissing interaction. Electrospray ionization mass spectrometry was used to detect kissing dimers in a multiequilibria mixture, whereas optical tweezers were applied to monitor the (un)folding trajectories of single RNA molecules. The experimental findings were rationalized by molecular dynamics simulations. Together, the results showed that the stacked adenines are indispensable for the tertiary interaction. By shielding the tertiary base pairs from solvent and reducing their fraying, the stacked adenines made terminal pairs act more like interior base pairs. The purine double-ring of adenine was essential for effective stacking, whereas additional functional groups modulated the stabilizing effects through varying hydrophobic and electrostatic forces. Furthermore, formation of the kissing complex was dominated by base pairing, whereas its dissociation was significantly influenced by the flanking bases. Together, these findings indicate that unpaired flanking nucleotides play essential roles in the formation of otherwise unstable two-base-pair RNA tertiary interactions.

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