PHOTOCHEMICAL & SPECTROSCOPIC PROBES OF RNA POLYMERASE

建议的工作是绘制新生的前端的路径 核糖核酸（RNA）穿过细菌酶RNA的表面 聚合酶和核糖体。 标记的RNA的原位制备 叠氮化物光亲和标记，以及随后的光解和分析 标记的产物，将显示酶的哪些亚基被 不断增长的核苷酸链和RNA链的长度 与DNA模板分离。 附着在RNA上的光谱探针， 其他网站将提供有关距离的补充信息， 大分子复合物上的位点，以及对 新生RNA的运动。 双功能螯合剂，类似物 乙二胺四乙酸，其可以共价连接到 生物分子，将被用作多用途光谱探针。 特别地，双功能螯合剂中的发光镧系离子 附着在新生RNA上的DNA将被用作能量转移的供体 问题研究 此外，从螯合的扩散增强的能量转移 溶液中的镧系元素将揭示 新生RNA和发色探针分子，在大分子 配合物 这些实验将提供新的分子知识， DNA转录和转录-翻译偶联机制; 更好地了解这些基本的生命过程可能最终会受益于 人体健康

The proposed work is to map the path of the leading end of nascent ribonucleic acid (RNA) across the surface of the bacterial enzyme RNA polymerase and on to the ribosome. In situ preparation of RNA tagged with azide photoaffinity labels, and subsequent photolysis and analysis of labeled products, will show which subunits of the enzyme are contacted by the growing nucleotide chain and also the RNA chain length at which RNA separates from DNA template. Spectroscopic probes attached to the RNA and other sites will provide complementary information about distances between sites on the macromolecular complex, and restrictions on the freedom of movement of nascent RNA. Bifunctional chelating agents, analogs of ethylenediaminetetraacetic acid which can be covalently attached to biological molecules, will be used as multipurpose spectroscopic probes. In particular, luminescent lanthanide ions in bifunctional chelating agents attached to nascent RNA will be used as donors for energy-transfer studies. Also, diffusion-enhanced energy transfer from chelated lanthanides in solution will reveal the accessibility of the leading end of nascent RNA, and of chromophoric probe molecules, in macromolecular complexes. These experiments will provide new knowledge of the molecular mechanisms of DNA transcription and transcription-translation coupling; better understanding of these basic life processes may ultimately benefit human health.