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Comparison of repair activity in different genomic regions

100%

Chakalova L., Russev G.

Acta Biochimica Polonica

1998

tom 45

nr 1

We have developed a quantitative technique to determine repair activity at defined genomic regions. Cells were treated with hydroxyurea to inhibit the replicative DNA synthesis and were incubated with 5-bromodeoxyuridine (BrdUrd) to label the regions undergoing repair. In the course of the labelling, the regions that were more actively repaired would incorporate more BrdUrd than the regions that were less actively repaired. Thus the kinetics of BrdUrd incorporation in the different sequences would reflect the kinetics of reparation of the respective regions. The total BrdUrd-containing, repaired DNA was isolated by immunoprecipitation with anti-BrdUrd antibody, and after controlled sonication, it was used as a template in quantitative PCR in which the amount of the product was directly proportional to the amount of template. This approach was used to address the question whether DNA repair after UV irradiation occurs in an uniformly random manner, or with preferences for certain regions. We found that, in Ehrlich ascites tumor cells, the repair efficiency was higher at the 5' end of the mouse β-globin domain than in the rest of the domain.

Effect of ethanol on structural transitions of DNA and polyphosphates under Ca2plus ions action in mixed solutions

67%

Hackl E. V., Blagoi Y. P.

Acta Biochimica Polonica

2000

tom 47

nr 1

In the present work using the IR spectroscopy method the effect of ethanol on structural transitions of DNA and polyphosphates under the action of Ca2+ ions in mixed solutions containing ethanol (0-25 vol.%) was studied. It was shown that, on its interaction with Ca2+ ions, in aqueous and mixed solutions DNA becomes transformed into compact form. With the increase of concentration of ethanol the degree of Ca2+-induced DNA compactisation rises. It was found that, in mixed solutions containing ethanol, Ca2+-induced DNA compactisation depends not only on the solution's dielectric permeability but also on the solution structure. On stabilisation of the water structure in the presence of low ethanol concentrations a stabilisation of the DNA macromolecule occurs that leads to the increase of the Ca2+ ion concentration necessary for DNA compactisation. Comparison of the effects of ethanol on Ca2+-induced structural transitions in DNA and polyphosphates in mixed solvents permits to suppose that at alcohol concentrations in solution resulting in disruption of the water spatial structure, some peculiarities are observed in the behavior of those molecules whose hydrophobic interactions are essential.

Ograniczanie wyników

2 Acta Biochimica Polonica

1 Blagoi Y. P.

1 Chakalova L.

1 Hackl E. V.

1 Russev G.

1 2000

1 1998

Wyniki wyszukiwania

Comparison of repair activity in different genomic regions

Effect of ethanol on structural transitions of DNA and polyphosphates under Ca2plus ions action in mixed solutions