RTE, a plant growth gene with a conserved role in metals

描述（由申请人提供）：广泛的长期目标是了解生物体控制过渡金属离子的机制以及这些金属（特别是铜）在细胞信号传导和调控途径中的作用。 铜稳态对人类健康至关重要，但细胞内铜运输的途径尚不清楚，并且已确定的铜传感和运输的成分相对较少。该提案重点关注 RTE，这是一种先前未描述的新序列基因，假设其编码铜的传感器、转运蛋白或伴侣。 RTE 基因高度保守，在动物中以单拷贝存在，在植物中以两个拷贝存在。该提案的目的是明确 RTE 在金属稳态中的作用。这些研究将主要集中在模式植物拟南芥的 RTE1 基因上。拟南芥遗传分析已将 RTE1 与乙烯信号转导中的两个铜结合蛋白联系起来。一种是需要铜的乙烯受体ETR1，其功能依赖于RTE1。另一个是 RAN1，它是人类 Menkes/Wilson 的 P 型 ATP 酶铜转运蛋白的同源物，它同样是乙烯受体功能所必需的。研究拟南芥中 RTE1 的这些连接的遗传和细胞基础将利用强大的遗传工具来了解 RTE 的细胞作用。 RTE 和 Menkes/Wilson 的同源物 CUA-1 的平行实验将在动物遗传模型秀丽隐杆线虫中进行，从而可以将动物系统的结果与植物系统的结果进行比较和整合。酿酒酵母和哺乳动物细胞培养物将分别用于乙烯结合测定和铜结合测定。拟议的实验将结合分子遗传学、细胞生物学和生化方法。 RTE 功能的阐明应该对理解金属稳态过程产生直接影响，从而造福人类健康。

DESCRIPTION (provided by applicant): The broad, long-term objective is to understand the mechanisms by which organisms control transition metal ions and the roles of these metals, particularly copper, in cell signaling and regulatory pathways. Copper homeostasis is of major importance to human health, yet the pathways for intracellular copper trafficking are unclear, and relatively few components in copper sensing and trafficking have been identified. This proposal focuses on RTE, a previously undescribed gene of novel sequence, which is hypothesized to encode a sensor, transporter or chaperone for copper. The RTE gene is highly conserved, and is present in a single copy in animals and in two copies in plants. The aim of this proposal is to define the role of RTE in metal homeostasis. The studies will center primarily on the RTE1 gene of the model plant Arabidopsis thaliana. Genetic analyses in Arabidopsis have linked RTE1 to two copper-binding proteins in ethylene signal transduction. One is the copper-requiring ethylene receptor ETR1, whose function is dependent on RTE1. The other is RAN1, a homolog of the human Menkes/Wilson's P-type ATPase copper transporter, which is likewise required for ethylene receptor function. Investigating the genetic and cellular basis for these connections to RTE1 in Arabidopsis will utilize powerful genetic tools for understanding the cellular roles of RTE. Parallel experiments with respect to RTE and the Menkes/Wilson's homolog CUA-1 will be performed in the animal genetic model Caenorhabditis elegans, allowing comparison and integration of results from an animal system with those in plants. Saccharomyces cerevisiae and mammalian cell culture will be employed for ethylene binding assays and copper binding assays, respectively. The proposed experiments will use a combination of molecular genetics, cell biology and biochemical methods. The elucidation of RTE function should have a direct impact on understanding the processes of metal homeostasis for the benefit of human health.