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July 1, 1966The Journal of General Physiology337 citationsOpen Access

Physical and Topological Properties of Circular DNA

JVJerome VinogradThe Wistar InstituteJLJacob LebowitzNational Institutes of Health

Key Points

  • This research investigates the physical and topological characteristics of various circular DNA molecules.
  • Characterization of different types of circular DNA, including single-stranded and duplex rings.
  • Experiments assessing sedimentation rates in alkaline solvents and thermal stability.
  • Analysis of the impact of single strand scission on properties of closed circular duplex DNA.
  • Circular DNA sediment rapidly in alkaline environments due to a topological unwinding barrier.
  • Duplex rings demonstrate greater thermodynamic stability in melting experiments compared to strand-separable DNA.
  • Single strand scission significantly alters closed circular duplex DNA, leading to formation of slower-sedimenting structures.

Abstract

Several types of circular DNA molecules are now known. These are classified as single-stranded rings, covalently closed duplex rings, and weakly bonded duplex rings containing an interruption in one or both strands. Single rings are exemplified by the viral DNA from phiX174 bacteriophage. Duplex rings appear to exist in a twisted configuration in neutral salt solutions at room temperature. Examples of such molecules are the DNA's from the papova group of tumor viruses and certain intracellular forms of phiX and lambda-DNA. These DNA's have several common properties which derive from the topological requirement that the winding number in such molecules is invariant. They sediment abnormally rapidly in alkaline (denaturing) solvents because of the topological barrier to unwinding. For the same basic reason these DNA's are thermodynamically more stable than the strand separable DNA's in thermal and alkaline melting experiments. The introduction of one single strand scission has a profound effect on the properties of closed circular duplex DNA's. In neutral solutions a scission appears to generate a swivel in the complementary strand at a site in the helix opposite to the scission. The twists are then released and a slower sedimenting, weakly closed circular duplex is formed. Such circular duplexes exhibit normal melting behavior, and in alkali dissociate to form circular and linear single strands which sediment at different velocities. Weakly closed circular duplexes containing an interruption in each strand are formed by intramolecular cyclization of viral lambda-DNA. A third kind of weakly closed circular duplex is formed by reannealing single strands derived from circularly permuted T2 DNA. These reconstituted duplexes again contain an interruption in each strand though not necessarily regularly spaced with respect to each other.

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Cite This Study

Vinograd et al. (1966) studied this question.

synapsesocial.com/papers/6a0ccba3291fe4aa62625576https://doi.org/10.1085/jgp.49.6.103
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