METALLOTHIONE EXPRESSION REGULATED BY OXIDATIVE STRESS

描述 利用小鼠肺上皮细胞和巨噬细胞系，申请人 提出了三个具体目标：1)表征Cd诱导的MT水平 蛋白质、MT mRNA和MT基因转录(通过瞬时转基因)，在 肺上皮细胞系与巨噬细胞系；2)识别顺式调控 负责Cd诱导转录的元件，重点关注Cd的作用 金属反应元件(MRE)和抗氧化反应元件 (Ares)；以及3)表征MRE的磷酸化状态 ~(32)P标记的镉或过氧化氢处理的转录因子(MTF) 细胞。这项研究将试图回答两个基本问题：1) 抗氧化反应元件在镉诱导MT基因中的重要作用 表情？2)Cd和H_2O_2是否都能激活金属硫蛋白 通过共同的途径转录--通过改变磷酸化 MTF的状况？拟议的研究旨在建立机械论 环境毒素Cd、金属诱导的ROS产生、 金属硫蛋白基因的表达与肺的恶化有关 疾病。

DESCRIPTION Using mouse lung epithelial and macrophage cell lines, the applicant proposes three specific aims: 1) characterize the levels of Cd-induced MT protein, MT mRNA, and MT gene transcription (via transient transfection), in lung epithelial vs. macrophage cell lines; 2) identify the cis regulatory elements responsible for Cd-induced transcription, focussing on the role of both metal response elements (MREs) and antioxidant response elements (AREs); and 3) characterize the phosphorylaton state of the MRE transcription factor (MTF) isolated from Cd- or H2O2-treated, 32P-labelled cells. The research will seek to answer two basic questions: 1) Does the antioxidant response element play an important role in Cd induced MT gene expression? and 2) Might both Cd and H2O2 activate metallothionein transcription through a common pathway -- by altering the phosphorylation state of MTF? The proposed research is designed to establish mechanistic linkages between the environmental toxin Cd, metal-induced ROS production, metallothionein gene expression, and the exacerbation of lung related diseases.