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：这是一个曾经修正的建议，继续博士Smulson的 为了实现这一目标，提出了三个目标的研究。 第一个是试图澄清的相互作用， 染色质中DNA断裂的PADPRP。初步研究将评估 在修复过程中PADPRP在DNA末端和末端的循环，使用 细菌表达的PADPRP突变蛋白。此外，稳定使用 转染的同步化3 T3-L1细胞，将寻求证据， 建立PADPRP与DNA复制/修复的直接作用， 定义与DNA聚合酶α的物理联系，以及 多酶DNA复制复合体。PADPRP的裂解 在该系统中的细胞凋亡过程中也将被评估。目标二是 设计用于确定聚（ADP-核糖）信号是否产生 随后的DNA链断裂发挥了重要作用， p53蛋白的积累和细胞停滞在G1期直至修复 可以继续了在目的III中，PADPRP在V（D）J中的机制作用 重组，其与链断裂共享若干酶特征 修复过程中，将进行探索。的影响 PADPRP表达构建体在这一过程中将被检查， 表达RAG 1和RAG 2基因的小鼠3 T3细胞， 通过使用新建立的“敲除”小鼠来对整个动物进行研究。