ALTERATION OF ERCC GENE EXPRESSION IN VITRO AND IN VIVO

描述：该项目的目标是确定是否改变 发生在不同核苷酸切除修复酶的表达中 在老化过程中。 我们的初步证据表明，至少有一个 这些DNA修复酶ERCC 3或BTF 2的p89 DNA解旋酶 转录复合物，在体外下调，而另一个 ERCC 1酶不是。 有趣的是，ERCC 3基因产物是 与科凯恩综合征有关，有些人认为这是一种 “过早衰老”综合征，而ERCC 1与 科凯恩氏病或着色性干皮病。 基础表达的改变 其他ERCC酶和其他几种与DNA相关的酶 修复和转录，将在体外和体内测定。 vivo. 体外研究包括监测人类二倍体 随着细胞的衰老。 体内研究包括比较 年轻人（20-30岁）和老年人（>70岁）之间的表达 人口。 这些酶对压力的反应能力 不同的DNA损伤剂诱导的DNA损伤也将进行比较， 年轻人和老年人的对比 由于ERCC 3/p89解旋酶是 只是BTPII复杂的5个组件之一，他还将监测 其他四个组成部分。 该项目的第二阶段将涉及 在人成纤维细胞中表达特异性ERCC/XP酶并确定 如果体外寿命被改变。 项目的最后阶段 包括分离ERCC 3的鼠同源物（已知的ERCC 3同源物）。 下调），然后产生小鼠 过表达 ERCC 3/p89解旋酶 以确定是否发生改变， 平均（50%）或最大（95%）寿命。

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.