DNA with damage in both strands as affinity probes and nucleotide excision repair substrates

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• 作者：N. V. Lukyanchikova ; I. O. Petruseva ; A. N. Evdokimov…
• 刊名：Biochemistry (Moscow)
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：81
• 期：3
• 页码：263-274
• 全文大小：1,032 KB
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• 作者单位：N. V. Lukyanchikova (1) (2)
  I. O. Petruseva (1)
  A. N. Evdokimov (1)
  V. N. Silnikov (1)
  O. I. Lavrik (1) (2) (3)
  
  1. Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090, Novosibirsk, Russia
  2. Novosibirsk State University, 630090, Novosibirsk, Russia
  3. Altai State University, 656049, Barnaul, Russia
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Biochemistry
  Bioorganic Chemistry
  Microbiology
  Biomedicine
  Russian Library of Science
  
• 出版者：MAIK Nauka/Interperiodica distributed exclusively by Springer Science+Business Media LLC.
• ISSN：1608-3040

文摘

Nucleotide excision repair (NER) is a multistep process of recognition and elimination of a wide spectrum of damages that cause significant distortions in DNA structure, such as UV-induced damage and bulky chemical adducts. A series of model DNAs containing new bulky fluoro-azidobenzoyl photoactive lesion dCFAB and well-recognized nonnucleoside lesions nFlu and nAnt have been designed and their interaction with repair proteins investigated. We demonstrate that modified DNA duplexes dCFAB/dG (probe I), dCFAB/nFlu₊₄ (probe II), and dCFAB/nFlu₋₃ (probe III) have increased (as compared to unmodified DNA, umDNA) structure-dependent affinity for XPC—HR23B (Kd _um > Kd _I > Kd _II ≈ Kd _III ) and differentially crosslink to XPC and proteins of NER-competent extracts. The presence of dCFAB results in (i) decreased melting temperature (ΔT_m = −3°C) and (ii) 12° DNA bending. The extended dCFAB/dG-DNA (137 bp) was demonstrated to be an effective NER substrate. Lack of correlation between the affinity to XPC—HR23B and substrate properties of the model DNA suggests a high impact of the verification stage on the overall NER process. In addition, DNAs containing closely positioned, well-recognized lesions in the complementary strands represent hardly repairable (dCFAB/nFlu₊₄, dCFAB/nFlu₋₃) or irreparable (nFlu/nFlu₊₄, nFlu/nFlu₋₃, nAnt/nFlu₊₄, nAnt/nFlu₋₃) structures. Our data provide evidence that the NER system of higher eukaryotes recognizes and eliminates damaged DNA fragments on a multi-criterion basis.