On the stability of double stranded nucleic acids.
We present the first pressure-versus-temperature phase diagram for the helix-to-coil transition of double stranded nucleic acids. The thermodynamic stability of a nucleic acid duplex is a complex function of temperature and pressure and strongly depends on the denaturation temperature, T(M), of the duplex at atmospheric pressure. Depending upon T(M), pressure, and temperature, the phase diagram shows that pressure may stabilize, destabilize, or have no effect on the conformational state of DNA. To verify the phase diagram, we have conducted high-pressure UV melting experiments on poly(dIdC)poly(dIdC), a DNA duplex, poly(rA)poly(rU), an RNA duplex, and poly(dA)poly(rU), a DNA/RNA hybrid duplex. The T(M) values of these duplexes have been modulated by altering the solution ionic strength. Significantly, at low salt, these three duplexes have helix-to-coil transition temperatures of 50 degrees C or less. In agreement with the derived phase diagram, we found that the polymeric duplexes were destabilized by pressure if the T(M) is < approximately 50 degrees C. However, these duplexes were stabilized by pressure if the T(M) is > approximately 50 degrees C. The DNA/RNA hybrid duplex, poly(dA)poly(rU), with a T(M) of 31 degrees C in 20 mM NaCl undergoes a pressure-induced helix-to-coil transition at room temperature. This is the first report of pressure-induced denaturation of a nucleic acid duplex and provides new insights into the molecular forces stabilizing these structures.
Dubins
D N
DN
Department of Pharmaceutical Sciences, Faculty of Pharmacy, University of Toronto, 19 Russell Street, Toronto, Ontario M5S 2S2, Canada.
Lee
A
A
Macgregor
R B
RB
Chalikian
T V
TV
D004247:DNA / Q000737:chemistry*; D006358:Hot Temperature; D009690:Nucleic Acid Conformation; D011061:Poly A / Q000737:chemistry; D011063:Poly A-U / Q000737:chemistry; D011072:Poly U / Q000737:chemistry; D011089:Polydeoxyribonucleotides / Q000737:chemistry; D011312:Pressure; D012313:RNA / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D013816:Thermodynamics
D011089:Polydeoxyribonucleotides; D011063:Poly A-U; D011061:Poly A; C015465:poly(dA); C031156:poly d(I-C); D011072:Poly U; D012313:RNA; D004247:DNA