Understanding ER chaperone-mediated RNR regulation

项目摘要 Bip/Kar2是一种普遍保守的分子伴侣，以内质网为基础，执行一种 细胞中的各种功能，包括新合成的和变性的蛋白质“客户”的蛋白质折叠 以及对末端错误折叠的蛋白质进行靶向降解。在酵母中，Kar2功能受辅助伴侣调节 如Sec63、Jem1和Scj1。虽然Scj1和Jem1似乎有一些多余的功能，但之前的研究 已经证明了缺乏Scj1或Jem1的细胞之间的表型差异。Jem1和Jem1的具体作用 SCJ1在激活Kar2和他们的特定客户组合方面仍未确定。 所有的生物都需要正确和准确的DNA复制才能生长和繁殖。监管不善 DNA复制可能导致细胞死亡或癌症。我们最近的研究发现了Scj1和Scj1的作用 Kar2在调节基因组完整性方面的作用。虽然内质网伴侣功能和基因组完整性是基本的细胞 进程，它们之间以前没有建立任何连接。 我们最近的研究表明，这种内质网伴侣-基因组完整性连接可能在 哺乳动物细胞失去ERdj1(哺乳动物Kar2的共同伴侣，Bip)使细胞对DNA复制敏感 抑制剂，如曲拉平和羟基脲。任何降低细胞内DNA复制速度的策略都可能 形成了新的抗癌疗法的基础。 在这项提案中，我们希望获得进一步的机械性洞察，了解ER监护人和合作伙伴是如何 伴侣控制着DNA的复制。我们建议同时使用分子生物学和最先进的质量 为了实现我们提案中目标的目标，我们采用了光谱分析技术来检测酵母菌和癌细胞。 这项工作的范围对与DNA复制相关的各种疾病具有广泛的影响 以及ER分子伴侣的功能，包括许多类型的癌症、病毒感染和疟疾。 好了！

Project Summary BiP/Kar2 is a universally conserved molecular chaperone based in the endoplasmic reticulum that performs a variety of functions in the cell including protein folding of both newly synthesized and denatured protein “clients” and targeted degradation of terminally misfolded proteins. In yeast, Kar2 function is regulated by co-chaperones such as Sec63, Jem1 and Scj1. While Scj1 and Jem1 appear to have some redundant functions, previous studies have demonstrated phenotypic differences between cells lacking Scj1 or Jem1. The specific roles of Jem1 and Scj1 in activating Kar2 and their particular client portfolio remains undetermined. All organisms require correct and accurate replication of DNA to grow and proliferate. Misregulation of DNA replication can result in either cell death or cancer. Our recent studies have uncovered a role for Scj1 and Kar2 in regulating genome integrity. While ER chaperone function and genome integrity are fundamental cellular processes, no connection between them has previously been established. Our recent studies suggest that this ER chaperone-genome integrity connection may be conserved in mammalian cells as loss of ERdj1 (co-chaperone of mammalian Kar2, BiP) sensitizes cells to DNA replication inhibitors such as triapine and hydroxyurea. Any strategy that lowers the rate of DNA replication in cells may form the basis of novel anticancer therapies. In this proposal, we expect to gain further mechanistic insight into how ER chaperones and co- chaperones control DNA replication. We propose to use both molecular biology and state-of-the-art mass spectrometric techniques in and yeast and cancer cells to achieve the aims of the objectives in our proposal. The scope of this work has broad implications for a variety of diseases associated with DNA replication and ER molecular chaperone function, including many types of cancer, viral infection and malaria. !