Abstract
We have examined the influence of centrin 2 (Cen2) on the interaction of nucleotide excision repair factors (XPC-HR23b, RPA, and XPA) with 48-mer DNA duplexes bearing the dUMP derivative 5-{3-[6-(carboxyamidofluores-ceinyl)amidocapromoyl]allyl}-2′-deoxyuridine-5′-monophosphate. The fluorescein residue linked to the nucleotide base imitates a bulky lesion of DNA. Cen2 stimulated the binding and increased the yield of DNA adducts with XPC-HR23b, a protein recognizing bulky damages in DNA. Stimulation of the binding was most pronounced in the presence of Mg2+ and demonstrated a bell-shaped dependence on Cen2 concentration. The addition of Cen2 changed the stoichiometry of RPA-DNA complexes and diminished the yield of RPA-DNA covalent crosslinks. We have shown that Cen2 influences the binding of RPA and XPA with DNA, which results in formation of additional DNA-protein complexes possibly including Cen2. We have also found some evidence of direct contacts between Cen2 and DNA. These results in concert with the literature data suggest that Cen2 can be a regulatory element in the nucleotide excision repair system.
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Abbreviations
- Cen2:
-
centrin 2
- Flu-dUMP:
-
5-{3-[6-(carboxyamidofluoresceinyl)amidocapromoyl]allyl}-2′-deoxyuridine-5′-monophosphate
- 5I-dUMP:
-
5-iodo-2′-deoxyuridine-5′-monophosphate
- NER:
-
nucleotide excision repair
- RPA:
-
replicative protein A
- XPA:
-
Xeroderma pigmentosum factor of complementation group A
- XPC-HR23b:
-
Xeroderma pigmentosum factor of complementation group C complexed with yeast protein Rad23 homolog
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Original Russian Text © Y. S. Krasikova, N. I. Rechkunova, E. A. Maltseva, C. T. Craescu, I. O. Petruseva, O. I. Lavrik, 2012, published in Biokhimiya, 2012, Vol. 77, No. 4, pp. 442–451.
Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM11-294, February 19, 2012.
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Krasikova, Y.S., Rechkunova, N.I., Maltseva, E.A. et al. Influence of centrin 2 on the interaction of nucleotide excision repair factors with damaged DNA. Biochemistry Moscow 77, 346–353 (2012). https://doi.org/10.1134/S0006297912040050
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DOI: https://doi.org/10.1134/S0006297912040050