Wheat germ 5S ribosomal RNA common arm fragment conformations observed by 1H and 31P nuclear magnetic resonance spectroscopy.
The nonexchangeable protons of the common arm fragment of wheat germ (Triticum aestivum) ribosomal 5S RNA have been observed by means of high-resolution 500-MHz 1H NMR spectroscopy in D2O solution. Although NMR studies on the exchangeable protons support the presence of two distinct solution structures of the common arm fragment (and of the same base-paired segment in intact 5S rRNA), only a single conformation is manifested in the 1H NMR behavior of all of the H6 and H5 pyrimidine and most of the H8/H2 purine protons under the same salt conditions. The nonexchangeable protons near the base-paired helix have been assigned by a sequential strategy. Conformational features such as the presence of a cytidine-uridine (C.U) pair at the loop-helix junction and base stacking into the hairpin loop are evaluated from nuclear Overhauser enhancement spectroscopy (NOESY) data. Double-quantum filtered correlation spectroscopy (DQF-COSY) experiments show that most of the 26 riboses are in the C3'-endo conformation. Finally, backbone conformational changes induced by Mg2+ and heating have been monitored by 31P NMR spectroscopy. Our results show that the common arm RNA segment can assume two conformations which produce distinguishably different NMR environments at the base-pair hydrogen-bond imino protons but not at nonexchangeable base or ribose proton or backbone phosphate sites.
Wu
J J
JJ
Department of Biochemistry, Ohio State University, Columbus 43210.
Marshall
A G
AG
D001482:Base Composition; D001483:Base Sequence; D006859:Hydrogen; D008274:Magnesium; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010758:Phosphorus; D012335:RNA, Ribosomal; D012341:RNA, Ribosomal, 5S; D014908:Triticum
D012335:RNA, Ribosomal; D012341:RNA, Ribosomal, 5S; D010758:Phosphorus; D006859:Hydrogen; D008274:Magnesium