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Title: Solution conformation of an RNA hairpin loop

Journal Article · · Biochemistry; (USA)
DOI:https://doi.org/10.1021/bi00469a026· OSTI ID:6169836

The hairpin conformation adopted by the RNA sequence {sup 5{prime}}GCGAUUUCUGACCGCC{sup 3{prime}} has been studied by one-and two-dimensional NMR spectroscopy. Exchangeable imino spectra in 60 mM Na{sup +} indicate that the hairpin has a stem of six base pairs (indicated by boldface type) and a loop of three nucleotides. NOESY spectra of nonexchangeable protons confirm the formation of the stem region. The duplex has an A-conformation and contains an A{center dot}C apposition; a G{center dot}U base pair closes the loop region. The stem nucleotides have C3{prime}-endo sugar conformations, as expected of an A-form duplex, whereas the three loop nucleotides adopt C2{prime}-endo sugar puckers. Stacking within the loop, C{sub 8} upon the sugar of U{sub 7}, stabilizes the structure. The pH dependence of both the exchangeable and nonexchangeable NMR spectra is consistent with the formation of an A{sup +}{center dot}C base pair, protonated at the N1 position of adenine. The stability of the hairpin was probed by using absorbance melting curves. The hairpin structure with the A{sup +}{center dot}C base pair is about +2 kcal/mol less stable in free energy at 37{degree}C than the hairpin formed with an A{center dot}U pair replacing the A{sup +}{center dot}C pair.

OSTI ID:
6169836
Journal Information:
Biochemistry; (USA), Vol. 29:17; ISSN 0006-2960
Country of Publication:
United States
Language:
English