Gendron D, Delbecchi L, Bourgaux-Ramoisy D, Bourgaux P
Department of Microbiology, The Medical School, Université de Sherbrooke, Québec, Canada.
J Virol. 1996 Jul;70(7):4748-60. doi: 10.1128/JVI.70.7.4748-4760.1996.
Previous work from this laboratory has indicated that intramolecular homologous recombination of polyomavirus (Py) DNA is dependent upon promoter structure or function. In this report, we demonstrate that Py DNA contains not two but three binding sites for transcription factor YY1, all located on the late side of viral origin of replication (ori) and the third well within the VP1 coding sequence. This third site (Y3), which may or may not play a role in transcription regulation, is immediately adjacent to a previously described recombination hot spot (S1/S2). We found that Py replicons carrying an altered Y3 site recombined in a manner suggesting partial inactivation of the S1/S hot spot. Point mutations precluding the binding of YY1 to Y3 in vitro depressed hot spot activity in vivo; however, of the two reciprocal products reflecting recombination at this spot, only that carrying the mutated Y3 site arose at a reduced rate. These results are interpreted in light of a model assuming that recombination occurs within a transcriptionally active viral chromatin tethered to the nuclear matrix by YY1.
该实验室之前的研究表明,多瘤病毒(Py)DNA的分子内同源重组依赖于启动子结构或功能。在本报告中,我们证明Py DNA含有三个而非两个转录因子YY1结合位点,它们均位于病毒DNA复制起点(ori)的晚期区域,第三个位点在VP1编码序列内。这个第三个位点(Y3)可能参与转录调控,也可能不参与,它紧邻之前描述的一个重组热点(S1/S2)。我们发现携带改变的Y3位点的Py复制子以一种表明S1/S热点部分失活的方式进行重组。在体外阻止YY1与Y3结合的点突变在体内降低了热点活性;然而,在反映该位点重组的两个相互产物中,只有携带突变Y3位点的产物出现的频率降低。这些结果根据一个模型来解释,该模型假定重组发生在由YY1 tethered至核基质的转录活跃的病毒染色质内。 (注:“tethered”此处可能有误,暂按原文翻译)