CARCINOGENS AND CHROMATIN STRUCTURE AND FUNCTION

致癌物和染色质结构与功能

• 批准号: 2653960
• 负责人: MARK E SMULSON
• 金额: $ 19.96万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-06-01 至 2000-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2653960
• 关键词: 3T3 cells; ADP ribosylation; DNA directed DNA polymerase; DNA repair; DNA replication; adenosine diphosphate; apoptosis; carcinogen testing; chemical carcinogen; chemical carcinogenesis; chemical structure function; chromatin; enzyme activity; genetic recombination; laboratory mouse; mutagen testing; nucleic acid structure; ribose; transcription factor; transfection

DESCRIPTION: This is a once-amended proposal to continue Dr. Smulson's studies into Three aims are proposed to pursue this objective. The first is structured to attempt to clarify the interaction of PADPRP with DNA breaks in chromatin. Initial studies will evaluate the cycling of PADPRP on and off ends of DNA during repair, using bacterially-expressed PADPRP mutant proteins. Also, using stably transfected synchronized 3T3-L1 cells, evidence will be sought to establish the direct role of PADPRP with DNA replication/repair by defining the physical association with DNA polymerase alpha, as well as a multi-enzyme DNA replicative complex. The cleavage of PADPRP during apoptosis in this system also will be evaluated. Aim II is designed to determine whether the poly(ADP-ribose) signal generated subsequent to DNA strand breaks plays a significant role in the accumulation of p53 protein and the arrest of cells at G1 until repair can proceed. In aim III the mechanistic role of PADPRP in V(D)J recombination, which shares several enzymatic features with strand break rejoining during repair, will be explored. The effects of the PADPRP expression constructs on this process will be examined in mouse 3T3 cells expressing the RAG 1 and RAG 2 genes and in the context of the whole animal by use of newly-established "knockout" mice.

描述：这是继续继续Smulson博士的提议 提出了三个目标的研究来追求这一目标。 第一个结构是为了阐明的相互作用 染色质中有DNA断裂的PADPRP。初步研究将评估 修复过程中DNA的PADPADPRP循环，使用 细菌表达的PADPRP突变蛋白。另外，使用稳定 转染的同步3T3-L1细胞，将寻求证据 通过通过DNA复制/修复来建立PADPRP的直接作用 还定义与DNA聚合酶α的物理关联 作为多酶DNA复制复合物。 PADPRP的裂解 在该系统的凋亡过程中，还将评估。 AIM II是 旨在确定是否生成聚（ADP-核糖）信号 在DNA之后断裂在 p53蛋白的积累和在G1上停滞直至修复 可以继续。在AIM III中，PADPRP在V（d）J中的机械作用 重组，该重组具有多种酶促特征与链断裂 将在维修过程中重新加入。效果 将在此过程中检查PADPRP表达构造 表达抹布1和抹布2基因的鼠标3T3细胞在上下文中 通过使用新建立的“淘汰”小鼠，整个动物中的整个动物。