POST-TRANSLATIONAL MODIFICATION OF THE REPLICATIVE DNA POLYMERASE ALPHA

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 先前的研究表明，DNA聚合酶α（Pol 1）在细胞周期中受到严格调控，Pol 1水平的改变会导致检查点控制功能障碍，S期DNA损伤修复减少和基因组不稳定。 这些观察结果表明，Pol 1可能在复制过程中严重参与DNA损伤检测，以及检查点控制。 关于翻译后修饰在细胞周期中调节Pol 1或响应DNA损伤中的作用知之甚少。 一个可能的可能性是Pol 1通过泛素化或SUMO化进行翻译后修饰。 支持这一观点的是观察到Pol 1在有丝分裂的中期-后期过渡期间迅速消失，这表明Pol 1经历了泛素化，这是其降解的信号;然而，候选的泛素连接酶目前尚不清楚。 由于这一领域是相对未研究的事实，我们建议使用酵母双杂交筛选，以帮助阐明候选酵母Pol 1结合伙伴，可能参与调节Pol 1在细胞周期或DNA损伤反应。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Previous work has shown that DNA polymerase alpha (Pol1) is tightly regulated during the cell-cycle, and that altered levels of Pol1 can lead to checkpoint control dysfunction, reduced S-phase DNA damage repair, and genomic instability. These observations suggest that Pol1 may be heavily involved in DNA damage detection during replication, as well as checkpoint control. Very little is known about the role that post-translational modifications play in regulating Pol1 during the cell cycle or in response to DNA damage. A likely possibility is that Pol1 undergoes post-translational modification via ubiquitylation or SUMOylation. In support of this idea is the observation that Pol1 rapidly disappears during the metaphase-anaphase transition during mitosis, suggesting that Pol1 undergoes ubiquitylation that signals its degradation; however, the candidate ubiquitin ligase is currently unknown. Due to the fact that this field is relatively unstudied, we propose to use the yeast two-hybrid screen to help elucidate candidate yeast Pol1 binding partners that may be involved in the regulation of Pol1 during the cell-cycle or DNA damage response.