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Nucleic Acids Research, 1994, Vol. 22, No. 9 1681-1686
© 1994

STRUCTURAL BIOLOGY

DNA stability at temperatures typical for hyperthermophiles

Evelyne Marguet and Patrick Forterre*

Institut de Génétique et Microbiologie CNRS URA 1354, Université Paris-Sud, 91 405, Orsay Cedex, France

*To whom correspondence should be addressed

Received January 4, 1994. Revised March 30, 1994. Accepted March 30, 1994.

We have studied the fate of covalently-closed circular DNA in the temperature range from 95 to 107°C. Super-coiled plasmid was not denatured up to the highest temperature tested. However, it was progressively transformed into open DNA by cleavage and then denatured. Thermodegradation was not dependent on the DNA supercoiling density. In particular, DNA made positively supercoiled by an archaeal reverse gyrase was not more resistant to depurination and thermodegradation than negatively supercoiled DNA. Thermodegradation was similar in aerobic or anaerobic conditions but strongly reduced in the presence of physiological concentrations of K+ or Mg2+. These results indicate that the major problem faced by covalently closed DNA in hyperthermophilic conditions is not thermodenaturation, but thermodegradation, and that intracellular salt concentration is important for stability of DNA primary structure. Our data suggest that reverse gyrase is not directly required to protect DNA against thermodegradation or thermodenaturation.


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