Unusual biphasic melting behaviour of Rhodotorula gracilis DNA

Ultraviolet, fluorescence and CD spectral analysis suggested unusual structural features of Rhodotorula gracilis ATCC 90950 DNA. R. gracilis DNA exhibited 13% hyperchromicity at 260 nm as against 26% shown by calf thymus DNA. The biphasic melting curve, one phase between 88-92 degrees C and the other between 92-97 degrees C, was attributed to different unwinding pattern of R. gracilis DNA as a function of rise in temperature. The binding affinity of ethidium bromide to R. gracilis DNA determined was almost the same as that of calf thymus DNA. Fluorescence spectra with rise in temperature showed decrease in the quanta of fluorescence intensity after transition temperature, suggesting the quenching due to variation in structure. The CD spectra of R. gracilis DNA did not resemble the spectra of any of the known DNA forms and it showed increase in the magnitude of negative band with rise in temperature suggesting a B-C transition. Disruption of intermolecular and higher order structures by sonication and salt concentration did not change this behaviour implicating the influence of sequence and base composition of R. gracilis DNA on thermal melting transition.