TELOMERASES AND TELOMERASE MOLECULAR BIOLOGY

端粒具有维持染色体的富含G的重复DNA序列 结束稳定。端粒酶是一种类似逆转录酶的酶。 维持端粒序列的核糖核蛋白。端粒酶是 在大多数肿瘤细胞中表达，而在正常细胞中不表达。 端粒酶和端粒是抗癌的理想肿瘤特异性靶点 治疗策略。我们的初步研究表明， 首先是端粒酶抑制剂。我们的研究还表明，端粒酶 抑制物可诱导端粒缩短、染色体末端融合和 最终细胞死亡。细胞已经显示出适应端粒酶的能力 抑制力。 这项建议的目的是为了阐明(L)的分子结构 端粒酶和为我们的抗癌提供了一个更明确的靶点 策略；2)确定细胞适应的机制(S) 端粒酶抑制剂。这些目标将通过测绘和 克隆端粒酶蛋白和RNA模板的基因。穿过 我们与L项目合作，纯化的端粒酶将用作 用于克隆的纯化的端粒酶RNA的来源。体细胞遗传学将 用于绘制端粒酶活性图谱。微分学的新技术 显示器将用于检测端粒酶后激活的mRNAs表达 在端粒酶前激活的细胞中不表达的细胞。这个 定位将与候选cdna克隆的分离相协调。 为蛋白质和RNA加速鉴定正当性 克隆人。将对RNA的cdna克隆进行筛选，以确定合适的 模板序列、二级结构和体外定向的能力 体内端粒酶活性。端粒酶蛋白的cDNA将是 表达是为了产生抗体。 抗体和DNA克隆将作为探针用于研究 端粒酶在程序3和程序4肿瘤标本中的表达。 这些研究的数据将被方案2用于结构 建模和具体的药物设计。潜在的端粒酶抑制剂，如 作为反义核糖核酸的寡核苷酸将由L计划进行研究。其机制 对端粒酶抑制剂的适应性也将进行研究。假设 关于药物代谢，端粒酶上调和端粒酶 将测试其特异性。

Telomeres have a G-rich repeated DNA sequence that maintains chromosome end stability. Telomerase is a reverse transcriptase-like ribonucleoprotein that maintains the telomere sequence. Telomerase is expressed in the majority of tumor cells but not in normal cells. Telomerase and telomeres are ideal tumor-specific targets for anti-cancer therapeutic strategies. Our preliminary studies show the discovery of the first telomerase inhibitors. Our studies also show that telomerase inhibitors induce telomere shortening, chromosome end fusion, and eventually cell death. Cells have shown the ability to adapt to telomerase inhibition. The goals of this proposal are to l) elucidate the molecular structure of telomerase and provide a more defined target for our anti-cancer strategies, and 2) determine the mechanism(s) of cellular adaptation to telomerase inhibitors. These goals will be accomplished by mapping and cloning the genes for the telomerase proteins and RNA template. Through our collaboration with Program l, purified telomerase will be used as a source of purified telomerase RNA for cloning. Somatic cell genetics will be used to map the telomerase activity. The new technique of Differential Display will be used to detect mRNAs expressed in posttelomerase-activated cells that are not expressed in pre-telomerase activated cells. The mapping will be coordinated with the isolation of candidate cDNA clones for the proteins and RNA to accelerate the identification of legitimate clones. The cDNA clones of the RNA will be screened for appropriate template sequence, secondary structure, and ability to direct in vitro and in vivo telomerase activity. The cDNAs for telomerase proteins will be expressed for production of antibodies. The antibodies and DNA clones will serve as probes for use in studies of telomerase expression in Program 3 and in tumor specimens from Program 4. The data from these studies will be used by Program 2 for structure modeling and specific drug design. Potential telomerase inhibitors, such as anti RNA oligonucleotides will be studied by Program l. The mechanism of adaptation to telomerase inhibitors will also be studied. Hypotheses concerning drug metabolism, telomerase up-regulation and telomerase specificity will be tested.