The surface properties of transfer RNA (tRNA) were analyzed using a poly(ethylene glycol)/dextran aqueous two-phase system (ATPS), where the surface net hydrophobicity (HFS) and the local hydrophobicity (LH) were evaluated based on the partition coefficient of tRNA in the ATPS. According to the evaluated HFS values, the surface of the tRNA molecule was hydrophilic at 20° -40 °C, and it became hydrophobic at 50° -80 °C because of the exposure of the intrinsic nucleobases of tRNA. In contrast, the LH values were found to be maximal at 20° -40 °C. The conformation of tRNA was investigated by Raman and circular dichroism (CD) spectroscopies, corroborating the results with the calculated prediction of its secondary structure (Mfold). It was shown that 66% of A-form structure existed at room temperature; the base stacking (θ265) was gradually decreased, and the A-form structure (θ208) was denatured along with a sigmoid curve against the temperature increase; the denatured secondary structures were observed above 50° C by Mfold prediction. The HFS value of the DNA duplex was found to be hydrophilic, compared to that of the single-stranded DNA, indicating that the exposure of nucleobases is a key factor of the hydrophobic properties of nucleotides. We conclude that the hydrophobic property of the tRNA surface was directly affected by its conformational transition.
The hammerhead ribozyme (HHR) is one of smallest catalytic RNAs, composed of a catalytic core and three stems; it undergoes self-cleavage in the presence of divalent magnesium ions (Mg(2+)) or other cations. It is hypothesized that the function and metabolism of RNAs might be regulated via interaction with lipid membranes in the prebiotic world. Using synthetic RNAs that model the core fragment of hammerhead ribozyme-like assembly (HHR-a), we investigated the enhancement of the self-cleavage reaction of HHR-a induced by the liposomes, both in the absence and presence of Mg(2+). The HHR-a activity was enhanced by 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)/1,2-dioleoyl-sn-glycero-3-phosphocholine (DPPC) = 8/2 liposome with Mg(2+), while other liposomes did not so significant. In the presence of DOPE/DPPC = 8/2 liposome, the HHR-a activity was observed without Mg(2+), revealed by the conformational change of the HHR inhibitor complex induced by the interaction with the liposome. The UV resonance Raman spectroscopy analysis investigated the interaction between lipid molecules and nucleobases, suggesting that the ethanolamine group of DOPE molecules are assumed to act as monovalent cations alternative to Mg(2+), depending on the liposome membrane characteristics.
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