TY - JOUR
T1 - Conformational change of single-stranded RNAs induced by liposome binding
AU - Suga, Keishi
AU - Tanabe, Tomoyuki
AU - Tomita, Hibiki
AU - Shimanouchi, Toshinori
AU - Umakoshi, Hiroshi
N1 - Funding Information:
Next Generation World-Leading Researchers (NEXT Program, GR066) from the Council for Science and Technology Policy, Cabinet Office and Japan Society for the Promotion of Science (JSPS). Ministry of Education, Science, Sports and Culture of Japan (MEXT), Grants-in-Aid for Scientific Research (Nos. 19656203, 19656220 and 20360350); Global COE program ‘Bio-Environmental Chemistry’ of JSPS. One of the authors (K.S.) acknowledges scholarships as JSPS and GCOE fellow. Funding for open access charge: NEXT, JSPS, MEXT.
Funding Information:
The fundamental concept of this study was supported by the Research Group of ‘Membrane Stress Biotechnology’ and the Sigma Multidisciplinary Research Laboratory Group (Graduate School of Engineering Science, Osaka University) ‘Membranomics’. We acknowledge Prof. Dr Ryoichi Kuboi (Executive Director of San Francisco Center, Osaka University) for his fruitful discussion and advice on this research.
PY - 2011/11
Y1 - 2011/11
N2 - The interaction between single-stranded RNAs and liposomes was studied using UV, Fourier Transform Infrared spectroscopy (FTIR) and Circular Dichroism spectroscopy (CD). The effect of the surface characteristics of liposomes, which were composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and modified with cholesterol (Ch) or 1,2-dioleoyl-3-trimethylammonium propane (DOTAP), on the liposome-RNA interaction was investigated. The fluorescence of 6-(p-toluidino)naphthalene-2-sulfonate (TNS) embedded in the liposome surface (ε=30-40) was decreased in the presence of tRNA, suggesting that single-stranded tRNA could bind onto the liposome. The dehydration of -PO2 - -, guanine (G) and cytosine (C) of tRNA molecules in the presence of liposomes suggested both an electrostatic interaction (phosphate backbone of tRNA and trimethylammonium group of POPC, DOTAP) and a hydrophobic interaction (guanine or cytosine of tRNA and aliphatic tail of lipid). The tRNA conformation on the liposome was determined by CD spectroscopy. POPC/Ch (70/30) maintained tRNA conformation without any denaturation, while POPC/DOTAP(70/30) drastically denatured it. The mRNA translation was evaluated in an Escherichia coli cell-free translation system. POPC/Ch(70/30) enhanced expression of green fluorescent protein (GFP) (116) while POPC/DOTAP(70/30) inhibited (37), suggesting that the conformation of RNAs was closely related to the translation efficiency. Therefore, single-stranded RNAs could bind to liposomal membranes through electrostatic and hydrophobic attraction, after which conformational changes were induced depending on the liposome characteristics.
AB - The interaction between single-stranded RNAs and liposomes was studied using UV, Fourier Transform Infrared spectroscopy (FTIR) and Circular Dichroism spectroscopy (CD). The effect of the surface characteristics of liposomes, which were composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and modified with cholesterol (Ch) or 1,2-dioleoyl-3-trimethylammonium propane (DOTAP), on the liposome-RNA interaction was investigated. The fluorescence of 6-(p-toluidino)naphthalene-2-sulfonate (TNS) embedded in the liposome surface (ε=30-40) was decreased in the presence of tRNA, suggesting that single-stranded tRNA could bind onto the liposome. The dehydration of -PO2 - -, guanine (G) and cytosine (C) of tRNA molecules in the presence of liposomes suggested both an electrostatic interaction (phosphate backbone of tRNA and trimethylammonium group of POPC, DOTAP) and a hydrophobic interaction (guanine or cytosine of tRNA and aliphatic tail of lipid). The tRNA conformation on the liposome was determined by CD spectroscopy. POPC/Ch (70/30) maintained tRNA conformation without any denaturation, while POPC/DOTAP(70/30) drastically denatured it. The mRNA translation was evaluated in an Escherichia coli cell-free translation system. POPC/Ch(70/30) enhanced expression of green fluorescent protein (GFP) (116) while POPC/DOTAP(70/30) inhibited (37), suggesting that the conformation of RNAs was closely related to the translation efficiency. Therefore, single-stranded RNAs could bind to liposomal membranes through electrostatic and hydrophobic attraction, after which conformational changes were induced depending on the liposome characteristics.
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U2 - 10.1093/nar/gkr568
DO - 10.1093/nar/gkr568
M3 - Article
C2 - 21785134
AN - SCOPUS:80455124110
SN - 0305-1048
VL - 39
SP - 8891
EP - 8900
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 20
ER -