MECHANISMS OF FIDELITY AND MUTAGENESIS

这个项目的广泛的长期目标是了解突变是如何 发生于致癌物修饰的DNA的复制。的目标是 这项拨款建议是为了阐明dna聚合酶之间的相互作用。 和DNA的小凹槽。具体目标是确定哪些是 DNA小沟中的位置与DNA聚合酶相互作用 在复制期间。这些相互作用将在 复制正常的DNA碱基和致癌物质6-甲基鸟嘌呤- 修改过的基地。观察到正确和错误之间的差异 复制将提供关于聚合酶用于 高保真复制DNA。该项目与健康相关的是 在致癌方面。正常DNA复制过程中发生的突变 而致癌物修饰的DNA可能会导致癌症。了解 突变形成的机制可能会让我们预测 化合物是诱变剂。这项研究的设计是合成DNA类似物 其中，小凹槽中的位置被改变，使得它们不能 参与氢键。改性剂的掺入动力学研究 与修饰的DNA相对的核苷酸三磷酸将被测量。大型 复制速度的降低将表明改变的站点是 与聚合酶发生氢键作用。

The broad long-term goal of this project is to understand how mutations occur from the replication of carcinogen-modified DNA. The objective of this grant proposal is to elucidate the interactions between DNA polymerase and the minor groove of DNA. The specific aims are to determine which sites in the minor groove of the DNA interact with the DNA polymerase during replication. These interactions will be examined during the replication of the normal DNA bases and theta6-methylguanine, a carcinogen- modified base. Differences observed between correct and incorrect replication will give clues as to the mechanisms that polymerases use to replicate DNA with high fidelity. The health-relatedness of the project is in carcinogenesis. Mutations that occur during replication of normal DNA and carcinogen-modified DNA can lead to cancer. Understanding the mechanism of the formation of mutations might alloW us to predict which compounds are mutagens. The research design is to synthesize DNA analogs in which sites in the minor groove are altered so that they can not participate in hydrogen bonds. The kinetics of incorporation of modified nucleotide triphosphates opposite modified DNA will be measured. Large reduction in rates of replication will indicate that the altered site was involved in a hydrogen bond with the polymerase.