INBRE-2 REGULATION OF COPPER HOMEOSTASIS IN YEAST

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 适当的铜稳态对生命至关重要，所有细胞都通过诱导转录程序来响应有毒的铜过量或铜饥饿，这些转录程序将导致储存多余的铜或根据需要摄取细胞外的铜。过量的自由铜离子会导致活性氧物种(ROS)的产生。ROS是包括DNA在内的细胞大分子损伤的主要来源。酵母CUP1基因编码一种铜结合的金属硫蛋白，这是在有毒铜水平存在时生存所必需的。CUP1的诱导需要铜敏感激活剂ACE1和染色质重构子SWI/SNF，而CUP1启动子受动态组蛋白乙酰化。在最初的铜反应后，CUP1的转录也会迅速关闭。这种自动调节是一个主动的过程，它不仅涉及ACE1的释放，而且还涉及SWI/SNF的持续存在。我们还鉴定了一个CUP1反义转录本RUF5，它在CUP1关闭过程中转录。本研究的目的是了解染色质结构和修饰是如何调控CUP1转录周期的，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.