ALTERATION OF ERCC GENE EXPRESSION IN VITRO AND IN VIVO

描述：该项目的目标是确定是否需要更改 发生在不同的核苷酸切除修复酶的表达中 在衰老过程中。我们的初步证据表明，至少有一名 这些DNA修复酶，ERCC3或BTF2的p89 DNA解旋酶 转录复合体，在体外下调，而另一个 ERCC1酶不是。有趣的是，ERCC3基因产物是 与Cockayne综合征有关，被一些人认为是一种 “早衰”综合征，而ERCC1与 Cockayne病或着色性干皮病。蛋白基础表达的改变 其他ERCC酶，以及其他几种与DNA相关的酶 修复和转录，将在体外和在 活着。体外研究包括监测人类二倍体。 随着细胞的衰老而产生的细胞。活体研究包括比较 年轻人(20-30岁)和老年人(70岁)之间的表情 人类人口。这些酶对压力的反应能力 不同的DNA损伤剂诱导的DNA损伤也将在 年轻人口与老年人口的对比。因为ERCC3/p89解旋酶是 只有BTFII建筑群的5个组成部分中的一个，他还将监测 其他4个组成部分。该项目的第二阶段将涉及 特异性ERCC/XP酶在人成纤维细胞中的表达及鉴定 如果在体外寿命被改变。该项目的最后阶段 涉及分离ERCC3的小鼠同源物(已知的是 被下调)，然后产生过度表达 ERCC3/p89解旋酶，以确定在 平均寿命(50%)或最高寿命(95%)。

DESCRIPTION: The goal of this project is to determine if alterations occur in expression of different nucleotide excision repair enzymes during aging. Our preliminary evidence indicates that at least one of these DNA repair enzymes, ERCC3 or the p89 DNA helicase of the BTF2 transcription complex, is down regulated in vitro whereas another ERCC1 enzyme is not. Interestingly, the ERCC3 gene product is implicated in Cockayne's syndrome, considered by some to be a "premature aging" syndrome, whereas ERCC1 is not associated with Cockayne's or Xeroderma pigmentosum. Alterations in basal expression of other ERCC enzymes, and several other enzymes associated with DNA repair and transcription, will be determined both in vitro and in vivo. The in vitro studies involve monitoring mortal human diploid cells as the cells senesce. The in vivo studies involve comparing expression between a younger (20-30 years) and an older (>70 years) human population. The ability of these enzymes to respond to stress induced by different DNA damaging agents will also be compared in a younger versus older population. As the ERCC3/p89 helicase is only 1 of 5 components of the BTFII complex, he will also monitor the other 4 components. The second phase of the project will involve expressing specific ERCC/XP enzymes in human fibroblasts and determining if in vitro life span is altered. The final phase of the project involves isolating the murine homologue of ERCC3 (the one known to be down regulated), then producing mice overexpressing the ERCC3/p89 helicase to determine if alterations occur in either the mean (50%) or the maximum (95%) life span.