SYNTHESIS OF 15N LABELED RNA FRAGMENTS

Project Details

Description

DESCRIPTION: The long-term goals of this proposal are to develop and
demonstrate synthetic strategies necessary for the synthesis and utilization
of specifically modified or labeled RNA fragments. Molecules that are
site-specifically labeled with 15N and 13C can provide unique and
model-independent insight into local RNA structure, including hydrogen
bonding, protonation, hydration, protection from hydration and stacking.
The utility of 15N specifically labeled nucleosides will be expanded by
introducing 13C into the C8 position of both purines and the C2 position of
adenine. The resulting coupling with 15N in only one of a pair of purines
will allow otherwise similar NMR resonances to be distinguished.
Furthermore, the 13C can provide independent information on base stacking.
The procedures developed for RNA synthesis using specific 15N and 13C labels
will be made more widely available by adapting them to the phosphoramidite
method. Since many of the simpler RNA motifs like non-canonical base pairs,
single bulges, and tetraloops have now been characterized by 15N NMR, the
new NMR work will focus primarily on more complex systems, such as internal
loops, large hairpin loops, pseudoknots, kissing loops, tetraloop receptors,
ribozymes, and protein-RNA complexes. The use of 15N NMR to define metal
ion binding to RNA, a crucial aspect of its structure and function, will be
explored.
StatusFinished
Effective start/end date1/1/945/31/02

Funding

  • National Institute of General Medical Sciences
  • National Institute of General Medical Sciences
  • National Institute of General Medical Sciences: $343,870.00
  • National Institute of General Medical Sciences
  • National Institute of General Medical Sciences
  • National Institute of General Medical Sciences
  • National Institute of General Medical Sciences
  • National Institute of General Medical Sciences: $196,143.00

ASJC

  • Spectroscopy

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