Melting and premelting phenomenon in DNA by laser Raman scattering

Abstract

Abstract Raman spectra of DNA from calf thymus DNA have been taken over a wide range of temperatures (25°–95°) in both D 2 O and H 2 O. A study of the temperature dependence of the Raman spectra shows that the temperature profiles of the intensities and frequencies of the various bands fall into four different categories: (1) base bands that show a reversible increase in intensity prior to the melting region, i.e., a definite premelting phenomenon; (2) base bands that show little or no temperature dependence; (3) deoxyribose‐phosphate backbone vibrations that show no temperature dependence up to the melting region, at which point large decreases in intensity occur; and (4) slow frequency changes in certain in‐plane vibrations of guanine and adenine due to deuteration of the C‐8 hydrogen of these purines in D 2 O. Certain Raman bands arising from each of the four bases, adenine, thymine, guanine, and cytosine have been found to undergo a gradual increase in intensity prior to the melting region at which point large, abrupt increases in intensity occur. The carbonyl stretching band of thymine, involved in the interbase hydrogen bonding actually undergoes both a gradual shift to a lower frequency as well as an increase in intensity. These changes provide evidence that some change in the geometry of the bases relative to each other begins to occur around 50°C, well below the melting region of 70°–85°C. From the spectra taken at various temperatures, the DNA appears to remain in the B conformation until the melting point is reached, at which time the DNA progresses into a disordered random‐coil form. No A‐form conformation is found either in the premelting or the melting region.