Effects of DNA-protein crosslink lesions on DNA replication and transcription

DNA-蛋白质交联（DPC）是非常庞大的基因组损伤，并且不能通过核苷酸切除修复有效地修复，这表明DPC将对细胞中的DNA复制和转录产生有害影响。在这项研究中，我们制备了模型DNA基板含有位点特异性DPC和评估其对DNA复制和转录的影响在体外。滞后链上的DPC抑制了E. coif，以DPC大小依赖的方式。相反，前导链上的DPC对DnaB的进展没有影响。我们还制备了含有DPC的DNA模板，用于T7 RNA聚合酶的体外转录研究。我们目前正在评估转录和非转录链中DPC对转录的影响。

DNA-protein crosslinks (DPCs) are extremely bulky genomic lesions and are not efficiently repaired by nucleotide excision repair, suggesting that DPCs will exert detrimental effects on DNA replication and transcription in cells. In this study, we have prepared model DNA substrates containing site-specific DPCs and assessed their effects on DNA replication and transcription in vitro. DPCs on the lagging strand inhibited the progression of DnaB, the replicative helicase of E. coif, in a DPC size-dependent manner. Conversely DPCs on the leading strand had no effects on the progression of DnaB. We also have prepared DNA templates containing DPCs for in vitro transcriptions studies with T7 RNA polymerase. We are currently assessing the effects of DPCs in transcribed and nontranscribed strands on transcription.