Experimental and computational studies of the G[UUCG]C RNA tetraloop.
In prokaryotic ribosomal RNAs, most UUCG tetraloops are closed by a C-G base-pair. However, this preference is greatly reduced in eukaryotic rRNA species where many UUCG tetraloops are closed by G-C base-pairs. Here, biophysical properties of the C[UUCG]G and G[UUCG]C tetraloops are compared, using experimental and computational methods. Thermal denaturation experiments are used to derive thermodynamic parameters for the wild-type G[UUCG]C tetraloop and variants containing single deoxy substitutions in the loop. A comparison with analogous experiments on the C[UUCG]G motif shows that the two RNA species exhibit similar patterns in response to the substitutions, suggesting that their loop structures are similar. This conclusion is supported by NMR data that suggest that the essential UUCG loop structure is maintained in both tetraloops. However, NMR results show that the G[UUCG]C loop structure is disrupted prior to melting of the stem; this behavior is in contrast to the two-state behavior of the C[UUCG]G molecule. Stochastic dynamics simulations using the GB/SA continuum solvation model, run as a function of temperature, show rare conformational transitions in several G[UUCG]C simulations. These results lead to the conclusion that substitution of a G-C for a C-G closing base-pair increases the intrinsic flexibility of the UUCG loop.
Williams
D J
DJ
Department of Biochemistry, Washington University School of Medicine, St Louis, MO 63110, USA.
Hall
K B
KB
D001419:Bacteria / Q000235:genetics; D020029:Base Pairing / Q000235:genetics; D001483:Base Sequence; D003198:Computer Simulation; D003855:Deoxyribose / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005057:Eukaryotic Cells; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D018583:Pliability; D011522:Protons; D020871:RNA Stability / Q000235:genetics; D012335:RNA, Ribosomal / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D012997:Solvents; D013269:Stochastic Processes; D013696:Temperature; D013816:Thermodynamics
D009841:Oligonucleotides; D011522:Protons; D012335:RNA, Ribosomal; D012997:Solvents; D003855:Deoxyribose