INBRE-2 REGULATION OF COPPER HOMEOSTASIS IN YEAST

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 适当的铜稳态对于生命至关重要，所有细胞都会通过诱导转录程序来应对有毒的铜过量或铜饥饿，从而根据需要封存过量的铜或吸收细胞外铜。过量的游离铜离子会导致活性氧 (ROS) 的产生。 ROS 是细胞大分子（包括 DNA）损伤的主要来源。酵母 CUP1 基因编码一种铜结合金属硫蛋白，这是在有毒铜水平存在下生存所必需的。 CUP1 诱导需要铜感应激活剂 Ace1 和染色质重塑剂 SWI/SNF，并且 CUP1 启动子会发生动态组蛋白乙酰化。 在最初的铜反应后，CUP1 转录也会迅速关闭。这种自动调节是一个主动过程，不仅涉及 Ace1 的释放，还涉及 SWI/SNF 的持续存在。我们还鉴定了 CUP1 反义转录本 RUF5，它在 CUP1 关闭期间转录。本研究的目的是了解染色质结构和修饰如何调节CUP1转录周期，并探讨RUF5转录本在CUP1表达中的作用。

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. Proper copper homeostasis is essential for life, and all cells respond to toxic copper excess or copper starvation by inducing transcriptional programs that will result in sequestration of excess copper or uptake of extracellular copper, as needed. Excess free copper ions result in generation of reactive oxygen species (ROS). ROS are a major source of damage to cellular macromolecules, including DNA. The yeast CUP1 gene encodes a copper-binding metallothionein that is required for survival in the presence of toxic copper levels. CUP1 induction requires the copper-sensing activator Ace1 and the chromatin remodeler SWI/SNF, and the CUP1 promoter is subject to dynamic histone acetylation. CUP1 transcription is also rapidly shut down after the initial copper response. This autoregulation is an active process, which involves not only release of Ace1 but also the continued presence of SWI/SNF. We have also identified a CUP1 antisense transcript, RUF5, which is transcribed during CUP1 shutdown. The goal of this study is to understand how chromatin structure and modification regulate the CUP1 transcription cycle, and to explore the role of the RUF5 transcript in CUP1 expression.