INTERACTIONS BETWEEN TELOMERIC DNA PRIMERS & TELOMERASE

端粒的完整性对染色体的稳定性至关重要， 最近发现，其他细胞功能，包括核 师.端粒序列的补充，逐渐失去了作为一个 常规DNA复制的结果，是由专门的 核糖核蛋白逆转录酶/端粒酶，它增加了端粒 通过复制内部RNA模板将DNA连接到DNA 3'末端。最近的数据 包括动力学研究表明，端粒酶与 引物序列5'到3'区域，参与与RNA的碱基配对 模板在引物识别中功能重要， 伸长过程中的易位。这种互动一直是 建议通过第二个（滞后产物）结合位点进行， 端粒酶在目前的“双位点”模型。这项提案将剖析 这种分子水平上的相互作用。具体目标是：（1） 探测端粒酶滞后产物位点之间的特异性接触， 和其DNA引物底物进行DNA足迹分析; DNA上的序列特异性官能团对 DNA引物与端粒酶滞后位点的相互作用 功能修饰干扰试验; 3）研究 端粒酶DNA在端粒酶与端粒DNA相互作用中的作用 引物这项研究将为端粒的生物合成和 一般来说，核酸的聚合。

The integrity of telomeres is essential for chromosomal stability and, it has been found recently, other cellular functions including nuclear division. The replenishment of telomeric sequences, lost gradually as a result of conventional DNA replication, is carried out by the specialized ribonucleoprotein reverse transcriptase/telomerase, which adds telomeric DNA to a DNA 3' end by copying an internal RNA template. Recent data including kinetic studies show that the interaction of telomerase with primer sequences 5' to the 3' region involved in base pairing to the RNA template is functionally important in primer recognition and translocation during processive elongation. This interaction has been proposed to be carried out by a second (lagging product) binding site of telomerase in the current 'two-site' models. This proposal will dissect this interaction at the molecular level. The specific aims are to 1) probe the specific contacts between the telomerase lagging product site and its DNA primer substrate by DNA footprinting analysis; 2) analyze the contribution of sequence-specific functional groups on DNA to the interaction of DNA primer with the lagging site of telomerase by functional modification interference assays; 3) study the role of the telomerase DNA in the interaction between telomerase and telomeric DNA primer. This study will provide insights into telomere biosynthesis and polymerization of nucleic acids in general.