DNA with Damage in Both Strands as Affinity Probes and Nucleotide Excision Repair Substrates.
作者： N. V. Lukyanchikova , I. O. Petruseva , A. N. Evdokimov , V. N. Silnikov , O. I. Lavrik
DOI: 10.1134/S0006297916030093
关键词:
摘要: Nucleotide excision repair (NER) is a multistep process of recognition and elimination wide spectrum damages that cause significant distortions in DNA structure, such as UV-induced damage bulky chemical adducts. A series model DNAs containing new fluoro-azidobenzoyl photoactive lesion dCFAB well-recognized nonnucleoside lesions nFlu nAnt have been designed their interaction with proteins investigated. We demonstrate modified duplexes dCFAB/dG (probe I), dCFAB/nFlu+4 II), dCFAB/nFlu−3 III) increased (as compared to unmodified DNA, umDNA) structure-dependent affinity for XPC—HR23B (Kd um > Kd I II ≈ III ) differentially crosslink XPC NER-competent extracts. The presence results (i) decreased melting temperature (ΔTm = −3°C) (ii) 12° bending. extended dCFAB/dG-DNA (137 bp) was demonstrated be an effective NER substrate. Lack correlation between the substrate properties suggests high impact verification stage on overall process. In addition, closely positioned, 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 system higher eukaryotes recognizes eliminates damaged fragments multi-criterion basis.
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