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Biomedical subjects

B M Sutherland

Publications and source records attributed to B M Sutherland.

At least 91 records · Page 5Linked to original sources
PubMed1975

The human leukocyte photoreactivating enzyme.

A photoreactivating enzyme from human leukocytes has been isolated and characterized. The enzyme requires DNA irradiated with ultraviolet light (220-300 nm) as substrate, and visible light (300-600 nm) for catalysis. In the reaction, the enzyme converts cyclobutyl pyrimidine dimers in the DNA to monomer pyrimidines. The enzyme has an apparent monomer molecular weight of 40,000 and tends to form aggregates. The pH optimum of 7.2 and absence of a requirement for metal ions are similar to the requirements of the Escherichia coli enzyme; however, the ionic strength optimum of 0.05 is much lower than those for other photoreactivating enzymes. The demonstration that human cells possess photoreactivating enzyme implies that a direct test by photoreactivation may be made of the role of pyrimidine dimers in the induction of abnormal cell growth.

Binding Sites↗
PubMed1975

Xeroderma pigmentosum cells contain low levels of photoreactivating enzyme.

Fibroblasts from patients with xeroderma pigmentosum contain low levels of photoreactivating enzyme in comparison to normal cells. Levels vary from 0 (line 1199) to 50 (line 1259) percent of normal. The depressed enzyme levels are not an artifact of low growth rate, age of cell donor, cell culture conditions, assay conditions, the presence of inhibitors, or mycoplasma contamination. We show that human fibroblasts can monomerize pyrimidine dimers in vivo.

Cell Line↗
PubMed1969

Mechanisms of inhibition of pyrimidine dimer formation in deoxyribonucleic acid by acridine dyes.

The ultraviolet (UV)-induced formation of cyclobutyl pyrimidine dimers in Escherichia coli deoxyribonucleic acid (DNA) in vitro has been investigated in terms of the mechanism of inhibition by acridine dyes, the effect on dimer yield of specific singlet and triplet quenchers, and the mechanism of dimer formation. Our results indicate that (a) energy transfer is important in dimer reduction by acridines, (b) this transfer occurs from the singlet (S(1)) of DNA, and (c) at room temperature triplet quenchers do not reduce dimer yield in DNA.

Acetone↗
PubMed1969

Inhibition of pyrimidine dimer formation in DNA by cationic molecules: role of energy transfer.

In addition to the acridine dyes, acridine orange and proflavine, we find that three other cationic molecules which bind to DNA-ethidium bromide, chloroquine, and methyl green-inhibit the production of cyclobutyl pyrimidine dimers by ultraviolet radiation. Intercalation is not necessary for dimer inhibition. The long range nature of the inhibition implies that energy transfer is responsible. The transfer is between the lowest excited singlet state of DNA and the acceptor singlet, and seems to involve the Förster mechanism.

Acridines↗
PubMed1969

Probes of DNA structure and interactions. Effects of copper II on ultraviolet-induced pyrimidine dimer formation.

Cu(II) affects the yield of cyclobutyl dimers induced in DNA by 254 nm radiation. The effects are a function of r, the ratio of Cu(II) to DNA phosphate, and of the ultraviolet (UV) fluence; they seem to reflect two types of copper complexes with DNA. The first probably involves "exterior" binding to the bases of native DNA and increases [unk]TT formation (without affecting [unk]UT yield) by raising the energy levels of bases other than thymine. The second seems to occur only at high ratios (rs) and only after the structure has been opened locally by UV radiation; it involves "interior" binding of Cu(II) to the bases. This complex tends to decrease dimer yield by holding the bases apart and/or by lowering the energy levels of bases other than thymine. These results illustrate the potential use of DNA photoproducts and ligands to probe the structure and interactions of DNA in vitro and perhaps also in vivo.

Carbon Isotopes↗
PubMed1968

Pyrimidine dimers in the DNA of Paramecium aurelia.

The production and fate of thymine-containing pyrimidine dimers in Paramecium aurelia DNA was investigated in three experimental series: production of dimers by UV irradiation, fate of dimers in the dark, and "loss of photoreactivability of dimers." It is shown that cyclobutyl dimers are made by UV irradiation of Paramecium DNA in vivo, that because of cytoplasmic absorption the number of dimers made in DNA irradiated in vivo is much lower than in DNA irradiated in vitro, that dimers are lost from animals incubated in the dark after irradiation, and that all the dimers that remain in the animals can be destroyed by photoreactivating illumination. Since mutation induction is photoreactivable, these and previous photoreactivation data suggest that pyrimidine dimers are important in mutation induction in P. aurelia.

Animals↗
PubMed1967

Photoreactivation in vivo of pyrimidine dimers in paramecium DNA.

Cells of Paramecium aurelia labeled with tritiated thymidine were irradiated with ultraviolet light and then were either exposed to photoreactivating light or kept in the dark as controls. In the controls, the level of thymine-containing pyrimidine dimers did not change, but in cells exposed to photoreactivating light such dimers were destroyed. This is the first demonstration in a eukaryote of in vivo photoreactivation of thymine-containing pyrimidine dimers.

Animals↗