Betaine can eliminate the base pair composition dependence of DNA melting.
We show that the amino acid analogue betaine shares with small tetraalkylammonium ions [Melchior, W. B., Jr., & von Hippel, P. H. (1973) Proc. Natl. Acad. Sci. U.S.A. 70, 298-302] the ability to reduce or even eliminate the base pair composition dependence of DNA thermal melting transitions. The "isostabilizing" concentration of betaine (at which AT and GC base pairs are equally stable) is approximately 5.2 M. Betaine exerts its isostabilizing effect without appreciably altering the conformation of double-stranded DNA from the B form. The presence of > 5 M betaine also does not greatly change the behavior of DNA as a polyelectrolyte; this lack of effect on electrostatic interactions is expected because betaine exists as a zwitterion near neutral pH. Study of DNA melting transitions in high concentrations of betaine thus allows the experimental separation of compositional and polyelectrolyte effects on DNA melting. As a consequence, betaine solutions can also be used to investigate DNA-protein interactions under isostabilizing (or close to isostabilizing) conditions, which has not been possible using isostabilizing salts. This potential is illustrated by examining the highly salt concentration-dependent interaction of ribonuclease A with DNA in concentrated betaine solutions.
Rees
W A
WA
Institute of Molecular Biology, University of Oregon, Eugene 97403.
Yager
T D
TD
Korte
J
J
von Hippel
P H
PH
D001482:Base Composition; D001622:Betaine / Q000494:pharmacology*; D004247:DNA / Q000737:chemistry* / Q000187:drug effects* / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000187:drug effects; D004563:Electrochemistry; D006358:Hot Temperature; D009690:Nucleic Acid Conformation; D011067:Poly dA-dT / Q000378:metabolism; D011189:Potassium Chloride / Q000494:pharmacology; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D013816:Thermodynamics
D004269:DNA, Bacterial; D011067:Poly dA-dT; D001622:Betaine; D011189:Potassium Chloride; D004247:DNA; D012259:Ribonuclease, Pancreatic