RNA and DNA interactions with zwitterionic and charged lipid membranes - a DSC and QCM-D study.
The aim of the present study is to establish under which conditions tRNA associates with phospholipid bilayers, and to explore how this interaction influences the lipid bilayer. For this purpose we have studied the association of tRNA or DNA of different sizes and degrees of base pairing with a set of model membrane systems with varying charge densities, composed of zwitterionic phosphatidylcholines (PC) in mixtures with anionic phosphatidylserine (PS) or cationic dioctadecyl-dimethyl-ammoniumbromide (DODAB), and with fluid or solid acyl-chains (oleoyl, myristoyl and palmitoyl). To prove and quantify the attractive interaction between tRNA and model-lipid membrane we used quartz crystal microbalance with dissipation (QCM-D) monitoring to study the tRNA adsorption to deposit phospholipid bilayers from solutions containing monovalent (Na(+)) or divalent (Ca(2+)) cations. The influence of the adsorbed polynucleic acids on the lipid phase transitions and lipid segregation was studied by means of differential scanning calorimetry (DSC). The basic findings are: i) tRNA adsorbs to zwitterionic liquid-crystalline and gel-phase phospholipid bilayers. The interaction is weak and reversible, and cannot be explained only on the basis of electrostatic attraction. ii) The adsorbed amount of tRNA is higher for liquid-crystalline bilayers compared to gel-phase bilayers, while the presence of divalent cations show no significant effect on the tRNA adsorption. iii) The adsorption of tRNA can lead to segregation in the mixed 1,2-dimyristoyl-sn-glycerol-3-phosphatidylcholine (DMPC)-1,2-dimyristoyl-sn-glycero-3-phosphatidylserine (DMPS) and DMPC-DODAB bilayers, where tRNA is likely excluded from the anionic DMPS-rich domains in the first system, and associated with the cationic DODAB-rich domains in the second system. iv) The addition of shorter polynucleic acids influence the chain melting transition and induce segregation in a mixed DMPC-DMPS system, while larger polynucleic acids do not influence the melting transition in these system. The results in this study on tRNA-phospholipid interactions can have implications for understanding its biological function in, e.g., the cell nuclei, as well as in applications in biotechnology and medicine.
Michanek
Agnes
A
Physical Chemistry 1, Lund University, P.O. Box 124, 22100 Lund, Sweden. agnes.zetterren@fkem1.lu.se
Kristen
Nora
N
Höök
Fredrik
F
Nylander
Tommy
T
Sparr
Emma
E
D000327:Adsorption; D002152:Calorimetry, Differential Scanning / Q000379:methods; D003460:Crystallization; D004247:DNA / Q000737:chemistry*; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry*; D008563:Membrane Lipids / Q000737:chemistry*; D055617:Micro-Electrical-Mechanical Systems / Q000379:methods; D010713:Phosphatidylcholines / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry; D011791:Quartz; D012313:RNA / Q000737:chemistry*; D012343:RNA, Transfer / Q000737:chemistry
D008051:Lipid Bilayers; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010743:Phospholipids; D011791:Quartz; D012313:RNA; D004247:DNA; D012343:RNA, Transfer; C017251:1,2-oleoylphosphatidylcholine; D004134:Dimyristoylphosphatidylcholine