Investigation of DNA structural changes by infrared spectroscopy

Abstract

AbstractDNA‐oriented samples of various origins were studied under different conditions of humidiity and sodium chloride content by means of infrared spectroscopy.(1) Oriented DNA (M. Lysodeikticus, E. coli, calf thymus and salmon sperm) films at 3–4% sodium chloride yield polarized spectra which show drastic changes at relative humidities (r.h.) between 94% and 0% indicative of conformational changes: B form → a form → disordered form The measurements of the infrared dichroism at frequencies of about 1230 cm−1 and at about 1090 cm−1 allow one to determine the orientation of the phosphate group, whereas the measurements at 1710 cm−1 characterize the base orientation. At humidities higher than 90% r.h. (B form) the bisector of OPO forms an angle of 70° relative to the helix axis, whereas at lower humidities, between 75% and 50% r.h. (A form) a rotation to about 45° is observed. Simultaneously, the 0—0 line of phosphate group changes its orientation from 55° to 65° to the helix when B → A transition takes place. The results are in general agreement with that of X‐ray diffraction and allow one to determine the orientation of the phosphate group with greater precision.(2) The B–A conformational change is not observed for satellite DNA, isolated from Cancer pagurus, of which the guanine + cytosine content is below 5%. As a function of decreasing humidities, one observes the transition: B form → disordered form A diagram of conformational changes of DNA's as a function of base composition and of r.h., suggests that B–A transition will occur for DNA of relatively higher G + C content, whereas for high (A + T) content, base sequence may be of importance.The B–A transition is prevented in DNA at a relatively high or very low sodium chloride content.