Reference - Detail
| RRC ID | 65011 |
|---|---|
| Author | Purohit NK, Robu M, Shah RG, Geacintov NE, Shah GM. |
| Title | Characterization of the interactions of PARP-1 with UV-damaged DNA in vivo and in vitro. |
| Journal | Sci Rep |
| Abstract |
The existing methodologies for studying robust responses of poly (ADP-ribose) polymerase-1 (PARP-1) to DNA damage with strand breaks are often not suitable for examining its subtle responses to altered DNA without strand breaks, such as UV-damaged DNA. Here we describe two novel assays with which we characterized the interaction of PARP-1 with UV-damaged DNA in vivo and in vitro. Using an in situ fractionation technique to selectively remove free PARP-1 while retaining the DNA-bound PARP-1, we demonstrate a direct recruitment of the endogenous or exogenous PARP-1 to the UV-lesion site in vivo after local irradiation. In addition, using the model oligonucleotides with single UV lesion surrounded by multiple restriction enzyme sites, we demonstrate in vitro that DDB2 and PARP-1 can simultaneously bind to UV-damaged DNA and that PARP-1 casts a bilateral asymmetric footprint from -12 to +9 nucleotides on either side of the UV-lesion. These techniques will permit characterization of different roles of PARP-1 in the repair of UV-damaged DNA and also allow the study of normal housekeeping roles of PARP-1 with undamaged DNA. |
| Volume | 6 |
| Pages | 19020 |
| Published | 2016-1-12 |
| DOI | 10.1038/srep19020 |
| PII | srep19020 |
| PMID | 26753915 |
| PMC | PMC4709520 |
| MeSH | Biocatalysis / radiation effects Chemical Precipitation DNA / metabolism DNA Damage* DNA Footprinting DNA Repair / radiation effects DNA-Binding Proteins / metabolism Deoxyribodipyrimidine Photo-Lyase / metabolism Green Fluorescent Proteins / metabolism Humans Models, Biological Poly(ADP-ribose) Polymerases / chemistry Poly(ADP-ribose) Polymerases / metabolism* Protein Binding / radiation effects Protein Domains Pyrimidine Dimers / metabolism Reproducibility of Results Ultraviolet Rays* |
| IF | 3.998 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 1 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | 10H(RCB0705) |