A spectroscopic and electron microscopic examination of the highly condensed DNA structures formed by denaturation in Mg(ClO 4) 2

Abstract

1. 1. Thermal denaturation in 1.5 M Mg(ClO 4) 2 of the DNA from bacteriophage λ results in four well-separated subtransitions, as monitored by the accompanying increase in absorbance. The midpoint of the hyperchromic spectrum is significantly lowered compared to either 1.5 M MgCl 2 or 3.0 M NaClO 4. 2. 2. The first two subtransitions are associated with the melting of the A · T-richest regions of the λ DNA, as revealed by electron micrographs following fixation with formaldehyde. 3. 3. Commencing with the third subtransition, an unusual DNA structure is observed in electron micrographs. In this structure the A · T-rich half of the molecule appears completely condensed, whereas the G · C-rich half remains native. 4. 4. During the fourth subtransition DNA molecules condense completely and eventually aggregate to form extremely high molecular weight particles containing centers of electron density. Tendrils of DNA, primarily duplex, radiate outward from these centers. 5. 5. The aggregation may be reversed by the removal of magnesium. The intramolecular condensation may be at least partly reversed by increasing the Mg(ClO 4) 2 concentrations to saturating levels.