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The cross talk between pathways in the repair of 8-oxo-7,8-dihydroguanine in mouse and human cells. | LitMetric

AI Article Synopsis

  • The study explores the repair of oxidatively damaged DNA, highlighting that both Base Excision Repair (BER) and Nucleotide Excision Repair (NER) pathways are involved, each contributing differently to the process.
  • Mouse embryo fibroblasts with specific genetic mutations related to DNA repair were exposed to potassium bromate to measure levels of a specific oxidized DNA base (8-oxoGua), revealing that BER, especially the OGG1 enzyme, plays a major role in its repair.
  • Findings suggest that the proteins CSB, XPC, and XPA operate in separate pathways, with XPA being newly identified as significant in managing oxidatively damaged DNA in human cells as well.

Article Abstract

Although oxidatively damaged DNA is repaired primarily via the base excision repair (BER) pathway, it is now evident that multiple subpathways are needed. Yet, their relative contributions and coordination are still unclear. Here, mouse embryo fibroblasts (MEFs) from selected nucleotide excision repair (NER) and/or BER mouse mutants with severe (Csb(m/m)/Xpa(-/-) and Csb(m/m)/Xpc(-/-)), mild (Csb(m/m)), or no progeria (Xpa(-/-), Xpc(-/-), Ogg1(-/-), Csb(m/m)/Ogg1(-/-)) or wild-type phenotype were exposed to an oxidizing agent, potassium bromate, and genomic 8-oxo-7,8-dihydroguanine (8-oxoGua) levels were measured by HPLC-ED. The same oxidized DNA base was measured in NER/BER-defective human cell lines obtained after transfection with replicative plasmids encoding siRNA targeting DNA repair genes. We show that both BER and NER factors contribute to the repair of 8-oxoGua, although to different extents, and that the repair profiles are similar in human compared to mouse cells. The BER DNA glycosylase OGG1 dominates 8-oxoGua repair, whereas NER (XPC, XPA) and transcription-coupled repair proteins (CSB and CSA) are similar, but minor contributors. The comparison of DNA oxidation levels in double versus single defective MEFs indicates increased oxidatively damaged DNA only when both CSB and XPC/XPA are defective, indicating that these proteins operate in different pathways. Moreover, we provide the first evidence of an involvement of XPA in the control of oxidatively damaged DNA in human primary cells.

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Source
http://dx.doi.org/10.1016/j.freeradbiomed.2012.08.593DOI Listing

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