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

DNA-蛋白质交联（DPC）是由各种试剂诱导的超大体积和普遍存在的DNA损伤。DPC可能会干扰DNA事务的许多方面。在本研究中，我们建立了在体外实验，以调查的影响DPC的真核转录的RNA聚合酶II催化，表明径流产品的形成是依赖于核细胞提取物的量和反应时间。然后，我们分析了模型大块病变（荧光素）对转录的影响。转录链上的黄绿素抑制转录，而非转录链上的黄绿素不抑制转录。我们还建立了一种新的方法，引入DPC之间的点击反应蛋白质叠氮化物和C5炔-dT。我们目前正在研究DPC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.