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• W2256512971 abstract "The biological effects due to inhibition by D/sub 2/O are discussed. The evidence of a primary isotope effect on DNA is as follows: instantaneous inhibition of cell division and DNA synthesis when cells are transferred into D/ sub 2/O; continued production of genetically defective sperm for a considerable period in deuterated male mice following return to regular drinking water, suggesting that D has made an impression on the information-bearing units; isolation of DNA in a modified form from deuterated anlmals; and the interference by D/sub 2/O with the genetically controlled synthesis of BETA -galactosidase in bacteria. The presence of D/sub 2/O greatly impairs the ability of the bacterial cell to respond to BETA -galactosides, but does not appreciably effect enzyme synthesis once it has been initiated. It is therefore postulated that the presence of heavy water modifies the functional state of DNA in vivo. A likely site of action may be on the keto-enol tautomerism of the purine and pyrimidine bases of DNA, since these equilibria result in rapid exchange of H (or D) with the medium. By an experimental study of this equilibrium, it was possible to calculate the percentate of keto form existing in solution at a particularmore » pH or pD. The presence of D/sub 2/O shifts the equilibrium toward stabilization of the keto form when comparison is made at the same pH and pD. The apparent pK of 8- thiognanine is 8.2 in H/sub 2/O and 8.6 in D/sub 2/O. Similarly, the pK of guanine was found to be 9.2 in H/sub 2/O and 10.0 in D/sub 2/O. The pK of thymine did not vary appreciably (10.0 in H/sub 2/O; 10.4 in D/sub 2/O). Stabilization of the keto tautomer of guanine by D/sub 2/O would be a very small effect at physiological pH since with a pK of 9.2 it would be predominantly in the keto form. The small magnitude of these isotope effects makes it unlikely that they would be significant in the metabolism of nucleotides and polynucleotides except possibly in the case of high molecular wt polynucleotides such as DNA, where thousands of stacked, H-bonded bases are involved. An investigation of the properties of DNA in aqueous and deuterated solutions showed no appreciable differences in the denaturation of DNA brought about by formamide (no appreciable differences were previously detected in the heat denaturation profiles of DNA or other polynucleotides in water or heavy water). Renaturation experiments showed an increased degree of renaturation (7-15% in DNA from chicken erythrocytes) in the presence of D/sub 2/O. This suggests that heat denaturation may be incomplete in D/sub 2/O, and that the persistence of a few H-bonded regions, by maintaining the strands in register, may promote helix reformation. It is proposed that the factors which lead to breakdown of DNA guanine-cytosine H- bonds may be dependent in part on shifts in the keto-enol equilibrium, and impairment of this mechanism by D/sub 2/O may result in incomplete strand separation. To explain the inhibitory effects on regulation of enzyme synthesis, it is suggested that genetic control of enzyme synthesis involves changes in the pattern of folding or the degree of H-bonding of DNA at speciffc regions which are predominantly guanine or cytosine in base sequence. (BBB)« less" @default.
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• W2256512971 date "1963-03-01" @default.
• W2256512971 modified "2023-09-23" @default.
• W2256512971 title "The use of deuterium isotope effects for studies of DNA structure and function." @default.
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