Open chromatin structures regulate the efficiencies of pre-RC formation and replication initiation in Epstein-Barr virus.
Open chromatin structures regulate the efficiencies of pre-RC formation and replication initiation in Epstein-Barr virus.
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DOI：
10.1083/jcb.201109105
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发表时间：
2012-08-20
期刊：
影响因子：
--
通讯作者：
Schepers A
中科院分区：
文献类型：
--
作者：
Papior P;Arteaga-Salas JM;Günther T;Grundhoff A;Schepers A
对EBV复制起源的研究表明，在基因组中灵活的mnase敏感位点形成了过量的复制前复合体。后生动物复制是否在随机或特定但灵活的位点开始是一个未解决的问题。起源识别复合体（ORC） DNA结合缺乏序列特异性，使基于基因组尺度染色质免疫沉淀（ChIP）的研究复杂化。爱泼斯坦-巴尔病毒（EBV）以染色质化的小染色体形式存在，并通过宿主复制机制进行复制。我们使用EBV在全基因组范围内研究了不同细胞周期阶段复制前复合体（pre-RC）组装区、复制起始区和微球菌核酸酶（MNase）敏感性之间的联系。EBV潜在起源的对称元（dyad symmetry element， DS）是一个建立良好且非常有效的预rc组装区域，它是EBV潜在起源的内部控制。我们确定了64个pre-RC区在空间上与57个短新生链（SNS）区相关。MNase实验显示，pre-RC和SNS区域与MNase敏感性增加的区域相关，这是来源强度的标志。有趣的是，虽然在空间上具有相关性，但pre-RC和SNS区域具有不同的特征。我们认为pre- rc是在灵活但不同的位点形成的，每个基因组和细胞周期只有少数被激活。
Studies of EBV replication origins demonstrate an excess of pre-replication complexes that are formed at flexible MNase-sensitive sites in the genome. Whether or not metazoan replication initiates at random or specific but flexible sites is an unsolved question. The lack of sequence specificity in origin recognition complex (ORC) DNA binding complicates genome-scale chromatin immunoprecipitation (ChIP)-based studies. Epstein-Barr virus (EBV) persists as chromatinized minichromosomes that are replicated by the host replication machinery. We used EBV to investigate the link between zones of pre-replication complex (pre-RC) assembly, replication initiation, and micrococcal nuclease (MNase) sensitivity at different cell cycle stages in a genome-wide fashion. The dyad symmetry element (DS) of EBV’s latent origin, a well-established and very efficient pre-RC assembly region, served as an internal control. We identified 64 pre-RC zones that correlate spatially with 57 short nascent strand (SNS) zones. MNase experiments revealed that pre-RC and SNS zones were linked to regions of increased MNase sensitivity, which is a marker of origin strength. Interestingly, although spatially correlated, pre-RC and SNS zones were characterized by different features. We propose that pre-RCs are formed at flexible but distinct sites, from which only a few are activated per single genome and cell cycle.
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影响因子：
6.7
作者：
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通讯作者：
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