Solid-phase synthesis and high-resolution NMR studies of two synthetic double-helical RNA dodecamers
- Univ. of Washington, Seattle (USA)
Ten-micromole solid-phase RNA synthesis has been successfully performed on an automated nucleic acid synthesizer with coupling efficiencies up to 99%, using the tert-butyldimethylsilyl group to protect the 2'-hydroxyl. The tert-butyldimethylsilyl group was easily removed by tetrabutylammonium fluoride under conditions in which virtually no 2'- to 3'-isomerization was found to occur. By use of this approach, the self-complementary RNA dodecamers r(CGCGAAUUCGCG) and r(CGCGUAUACGCG) were synthesized on an automated nucleic acid synthesizer, purified by TLC, and studied by high-resolution NMR. Imino protons were assigned from one-dimensional nuclear Overhauser effects. The nonexchangeable base, H1', and H2' protons were assigned by the sequential NOESY connectivity method. The NOE data from these two oligomers were analyzed qualitatively and compared to the ideal A- and B-type helix models of Arnott et al. (1972a,b). The internucleotide H6/H8 NOEs to the preceding H1' in r(CGCGUAUACGCG) were found to be sequence-dependent and probably reflect the roll angles between adjacent bases. The internucleotide H6/H8 to H2' NOEs of these oligomers correspond very well to an A-type conformation, but the interstrand adenine H2 NOEs to the following H1' were much stronger than those predicted from the fiber model. These strong interstrand NOEs can be rationalized by base pair slide to favor more interstrand base overlap.
- OSTI ID:
- 5476796
- Journal Information:
- Biochemistry; (USA), Vol. 28:6; ISSN 0006-2960
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
RNA
NUCLEAR MAGNETIC RESONANCE
ADENINES
BIOSYNTHESIS
CHEMICAL SHIFT
MOLECULAR STRUCTURE
OLIGONUCLEOTIDES
OVERHAUSER EFFECT
PROTONS
THIN-LAYER CHROMATOGRAPHY
AMINES
ANTIMETABOLITES
AROMATICS
AZAARENES
BARYONS
CHROMATOGRAPHY
DRUGS
ELEMENTARY PARTICLES
FERMIONS
HADRONS
HETEROCYCLIC COMPOUNDS
MAGNETIC RESONANCE
NUCLEIC ACIDS
NUCLEONS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PURINES
RESONANCE
SEPARATION PROCESSES
SYNTHESIS
550601* - Medicine- Unsealed Radionuclides in Diagnostics