Cationic lipids and cationic ligands induce DNA helix denaturation: detection of single stranded regions by KMnO4 probing.
Cationic lipids and cationic polymers are widely used in gene delivery. Using 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) as a cationic lipid, we have investigated the stability of the DNA in DOTAP:DNA complexes by probing with potassium permanganate (KMnO4). Interestingly, thymidines followed by a purine showed higher susceptibility to cationic ligand-mediated melting. Similar studies performed with other water-soluble cationic ligands such as polylysine, protamine sulfate and polyethyleneimine also demonstrated melting of the DNA but with variations. Small cations such as spermine and spermidine and a cationic detergent, cetyl trimethylammonium bromide, also rendered the DNA susceptible to modification by KMnO4. The data presented here provide direct proof for melting of DNA upon interaction with cationic lipids. Structural changes subsequent to binding of cationic lipids/ligands to DNA may lead to instability and formation of DNA bubbles in double-stranded DNA.
Prasad
T K
TK
Centre for Cellular and Molecular Biology, Uppal Road, 500 007 Hyderabad, India.
Gopal
Vijaya
V
Rao
N Madhusudhana
NM
D001483:Base Sequence; D002412:Cations; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D004277:DNA, Single-Stranded / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005229:Fatty Acids, Monounsaturated; D066298:In Vitro Techniques; D008024:Ligands; D008055:Lipids; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D011196:Potassium Permanganate; D011401:Promoter Regions, Genetic; D000644:Quaternary Ammonium Compounds
D002412:Cations; D015342:DNA Probes; D004277:DNA, Single-Stranded; D005229:Fatty Acids, Monounsaturated; D008024:Ligands; D008055:Lipids; D000644:Quaternary Ammonium Compounds; D011196:Potassium Permanganate; D004247:DNA; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane