REGULATION OF ANTIOXIDATIVE GENES AND AGING

描述（改编自申请人的摘要）： 本提案的目的是阐明监管机制， 控制铜锌超氧化物歧化酶（SOD）的表达， 过氧化氢酶，使用果蝇作为模型系统。 超氧 歧化酶和过氧化氢酶是抗氧化酶，它们协同作用， 除去H2O2。 这些酶的保护作用至关重要，因为 过氧化氢，在游离Fe的存在下，可以容易地导致 羟基自由基的产生，被认为是一个主要的罪魁祸首， 生物大分子氧化损伤的原因。 积累 氧化损伤的作用被认为在 各种疾病状态，包括癌症和动脉粥样硬化，以及衰老。 事实上，CuZn SOD与过氧化氢酶联合过表达， 被证明可以减少氧化分子损伤，延长寿命， 果蝇转基因。 本提案的前两个具体目标旨在测试 假设抗氧化基因表达受多种因素影响 监管水平。 在具体目标1中， 将测定过氧化氢酶表达和CuZn SOD表达 通过组织化学分析（使用β-半乳糖苷酶报告基因 基因）和/或免疫定位。 这将由第二个补充 具体目的是研究过氧化氢酶和铜锌的诱导作用， SOD，无论是对氧化应激方案的反应，还是对 荷尔蒙信号。 具体目标3的目的是确定 顺式作用调控元件，对特定模式至关重要 过氧化氢酶和CuZn SOD的表达。 为实现这些目标将 将修饰的启动子序列与报告基因融合并表征 果蝇转基因的表达模式。 在具体目标4中， 驱动异常表达的修饰的启动子将融合到 对应的天然基因结构域。 通过将这些转基因与 改变的启动子与未修饰的抗氧化转基因一起， 应该可以鉴定顺式调节元件， 这对延长寿命至关重要，因此为隔离铺平了道路 关键的反式调节因子。 这些研究最终可以为制定有效的策略提供参考 用于调节衰老的速度，然后可以应用于哺乳动物 系统.

DESCRIPTION (Adapted from the Applicant's Abstract): The long-term objective of this proposal is to elucidate the regulatory mechanisms governing the expression of CuZn superoxide dismutase (SOD) and catalase, using Drosophila melanogaster as a model system. Superoxide dismutase and catalase are antioxidative enzymes which act in tandem to remove H2O2. The protective role of these enzymes is crucial, because hydrogen peroxide, in the presence of free Fe, can readily lead to the generation of hydroxyl free radicals, thought to be a major culprit in the causation of oxidative damage to biomacromolecules. The accumulation of oxidative damage has been postulated to play a significant role in various disease states, including cancer and atherosclerosis, and aging. Indeed, the overexpression of CuZn SOD in conjunction with catalase has been shown to reduce oxidative molecular damage and extend life span in Drosophila transgenics. The first two Specific Aims of this proposal are designed to test the hypothesis that antioxidative gene expression is subject to multiple levels of regulation. In Specific Aim 1, the temporal and spatial expression of both catalase and CuZn SOD expression will be determined by means of histochemical analysis (using the b-galactosidase reporter gene) and/or immunolocalization. This will be complemented by the second Specific Aim, which is to examine the inducibility of catalase and CuZn SOD, either in response to regimens of oxidative stress or in response to hormonal signals. The objective of Specific Aim 3 is to identify cis-acting regulatory elements that are critical for specific patterns of catalase and CuZn SOD expression in vivo. This will be achieved by fusing modified promoter sequence to reporter genes and characterizing expression patterns in Drosophila transgenics. In specific Aim 4, modified promoters which drive aberrant expression will be fused to the corresponding native gene domains. By placing these transgenes with altered promoters together with non-modified antioxidative transgenes, it should be possible to identify cis-regulatory elements that are essential for life span extension and thus pave the way for isolation of critical trans-acting regulatory factors. Ultimately these studies may lease to development of effective strategies for modulating the rate of aging, which can then be applied to mammalian systems.