MECHANISMS OF METAL MUTAGENESIS: CR, NI, & BE

金属诱变机制：CR、NI、

• 批准号: 3458490
• 负责人: ELIZABETH T SNOW
• 金额: $ 10.92万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-06 至 1992-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3458490
• 关键词: DNA binding protein; DNA directed DNA polymerase; Escherichia coli; beryllium; cancer risk; cations; chemical carcinogen; chemical carcinogenesis; chromium; endonuclease; gene mutation; glycoproteins; hamsters; magnesium; metal metabolism; metalloproteins; metals; neoplasm /cancer epidemiology; nickel; nucleic acid probes; simian virus 40; tissue /cell culture; virus protein

A number of industrially important metals and carcinogens. Nickel and chromium stand out because they are human and animal carcinogens and also positive in short-term toxicity tests. Beryllium, an animal carcinogen and mutagen, is also positive in short-term tests. The molecular mechanisms by which these three metals act are not known. These metal-DNA interactions will be examined in a series of in vitro and in vivo experiments designed to measure the mechanisms and importance of metals in mutagenesis, DNA damage, polymerase function, and DNA-protein crosslinking. Metal mutagenesis, will be assayed in vitro with the M13mp2 forward-mutation assay using eukaryotic DNA polymerase-alpha. Mutations produced in vitro by the interactions of Be, Ni, or Cr will be quantitated, sequenced, and compared. The metal-induced mutations will be compared with the types of metal-induced DNA damage; base- and/or sequence-specifically will be assessed. Metal-induced nonspecific DNA damage also will be identified as DNA polymerase pause sites. Because DNA polymerase require divalent metal cations for activity and can be adversely affected by the wrong cations, the kinetics of DNA polymerase activity will be determined in the presence of the metal ions using kinetic primer-extension techniques. Since both Ni and Cr produce DNA- protein crosslinking in vitro, the effects of crosslinking on polymerase activity and mutagenesis will be studied using nonsequence-specific DNA binding proteins. Each metal's mutation spectrum will also be analyzed in vivo using an integrated shuttle-vector. Comparison of metal mutagenesis in vivo with the damage and mutation spectrum in vitro will provide evidence of whether the mechanisms are the same or not.

一些工业上重要的金属和致癌物质。 镍和铬之所以突出，是因为它们是人类的， 动物致癌物，在短期毒性试验中也呈阳性。 动物致癌物和诱变剂Berglycol也呈阳性， 短期测试。 这些分子机制 三种金属的行为是未知的。 这些金属-DNA相互作用将在一系列的研究中进行研究。 体外和体内实验旨在测量 金属在DNA诱变中的作用机制和重要性 损伤、聚合酶功能和DNA-蛋白质交联。 将使用M13 mp2在体外测定金属诱变 使用真核DNA聚合酶-α的正向突变测定。 Be、Ni或Cr相互作用在体外产生的突变 将被定量测序并进行比较 金属诱导 突变将与金属诱导的DNA类型进行比较， 损伤;将评估碱基特异性和/或序列特异性。 金属诱导的非特异性DNA损伤也将被确定为 DNA聚合酶暂停位点。 因为DNA聚合酶需要 二价金属阳离子的活性，并可能产生不利影响 DNA聚合酶活性的动力学 将在金属离子存在下使用动力学方法测定 引物延伸技术。 既然镍和铬都能产生DNA- 蛋白质体外交联，交联对 聚合酶活性和诱变将使用 非序列特异性DNA结合蛋白。 每种金属 突变谱也将在体内使用 整合穿梭载体。 金属诱变的比较 体内与体外损伤和突变谱将提供 是否相同的机制的证据。