Effects of DNA-protein cross-link lesions on eukaryotic transcription and replication

DNA-蛋白交联（DPCs）是一种由多种药物引起的超大体积且普遍存在的DNA损伤。dpc可能会干扰DNA交易的许多方面。在本研究中，我们通过体外实验研究了dpcs对RNA聚合酶II催化的真核生物转录的影响，结果表明，径流产物的形成取决于核细胞提取物的量和反应时间。然后，我们分析了模型大体积病变（荧光素）对转录的影响。荧光素在转录链上而不在非转录链上抑制转录。我们还建立了一种利用蛋白质叠氮化物与C5炔- dt之间的Click反应来引入DPCs的新方法。我们目前正在研究DPCs对RNA聚合酶II转录的影响。

DNA-protein cross-links (DPCs) are superbulky and ubiquitous DNA lesions induced by various agents. It is likely that DPCs interfere with many aspects of DNA transactions. In the present study, we established in vitroassays to investigate the effect ofDPCs on eukaryotic transcription catalyzed by RNA polymerase II, showing that the formation of runoff products are dependent on the amount of nuclear cell extracts and reaction time. We then analyzed the effect of a model bulky lesion (fluorescein) on transcription. Fluorescein on the transcribed strand but not on the nontrascribed strand inhibited the transcription. We also established a novel method to introduce DPCs by employing Click reactions between protein azides and C5 alkyne-dT. We are currently studying the effect of DPCs on transcription by RNA polymerase II.