Proceedings of the National Academy of Sciences · 1964 · 712 citations · 14 references
Ultraviolet LightChromatinThymine DimersBiochemistryPhotochemistryNatural SciencesPhotocarcinogenesisPhotobiologyBiotechnologyMolecular BiologyDna ReplicationPhototoxicityOligonucleotideIncubation Thymine DimersUv-c IrradiationPhotoprotectionUv Light IrradiationHealth Sciences
The study compares two E. coli K‑12 strains, AB1157 and its uv‑sensitive mutant AB1886, which carries a uvrA locus mutation. Cells were grown in H3‑thymidine, irradiated with UV, incubated in enriched minimal medium, then DNA was extracted with cold trichloroacetic acid, hydrolyzed in hot acid, and the resulting fractions were separated by paper chromatography. Thymine dimers were initially found in the acid‑insoluble fractions of both strains, but during incubation they were released into the acid‑soluble fraction only in AB1157, indicating excision by the uvr+ strain and suggesting enzymatic removal and DNA reconstruction preserve genetic integrity.
The fate of thymine dimers in DNA during incubation after uv light irradiation was studied in two strains of E. coli K-12. One was a multiply auxotrophic strain, AB1157, and the other was a uv-sensitive mutant, AB1886, derived from it. Strain AB1886 is unable to reactivate uv-irradiated Tl phage and is known to have a mutation at the uvrA locus. Cells were labeled in their DNA by growth with H/sup 3/thymidine, exposed to uv light, incubated in enriched minimal medium, and extracted with cold trichloroacetic acid. The acid precipitate and soluble fractions were hydrolyzed in hot acid, and the products were separated by paper chromatography. Thymine dimers were identified in the acid-insoluble fractions from both strains before incubation. During incubation thymine dimers were released into the acid-soluble fraction in the parental strain, AB1157, but not in the uv-sensitive strain AB1886. It is concluded that thymine dimers are excised from the DNA during the reactivation process in the uvr/sup +/ strain and that the sensitive uvr/sup -/ strain cannot do this. These findings suggest that the enzymatic removal of injured bases, including thymine dimers, and the reconstruction of the DNA from information on the complementary strand may be an important biologicalmore » mechanism for the preservation of DNA. (auth)« less
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