MOLECULAR PHARMACOLOGY OF TUMOR AND VIRUS INHIBITORS

肿瘤和病毒抑制剂的分子药理学

• 批准号: 6341823
• 负责人: Arthur Patrick Grollman
• 金额: $ 43.91万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-01-01 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6341823
• 关键词: DNA damage; DNA directed DNA polymerase; DNA repair; antiviral agents; carcinogenesis; carcinogenesis inhibitor; chemical binding; endonuclease; enzyme mechanism; enzyme substrate; frameshift mutation; gene mutation; molecular genetics; neoplasm /cancer genetics; neoplasm /cancer pharmacology; nucleic acid sequence; nucleic acid structure; oligonucleotides; site directed mutagenesis

This research focusses on the biological consequences of DNA damage with the overall goal of elucidating primary events in carcinogenesis. A principal theme is to establish relationships between the structure of damaged DNA and the functions of enzymes involved in DNA replication and repair. Novel experimental systems have been developed for this purpose. Site specific mutagenesis involves a strategy in which a shuttle plasmid vector, containing a single defined lesion, is allowed to replicate in mammalian cells or bacteria. The position of mutations induced is established by DNA sequence analysis. Primer-extension reactions, catalyzed by DNA polymerase, coupled with steady-state kinetic analysis, are used to explore translesional synthesis and mutagenic events in vitro. Our specific aims are (a) to establish models for frameshift mutagenesis in terms of misaligned DNA templates and kinetics governing translesional synthesis; (b) to elucidate the molecular basis underlying sequence context effects on base substitutions and deletions; (c) to understand the role of SOS functions in translesional synthesis; (d) to discover pathways by which mutations are generated during repair of bistrand abasic sites in DNA; (e) to demonstrates differences between DNA polymerases in their abilities to generate mutations arising from DNA damage; (f) to develop in vitro assays that predict mutagenic specificity for defined DNA lesions in vivo; (g) to explore mechanisms by which DNA damage enhances the frequency of homologous recombination in mammalian cells and bacteria; and (h) to establish the solution structure of misaligned intermediates formed during deletion mutagenesis. Additional studies are designed (a) to determined the substrate specificity of Fpg protein; (b) to establish the role of the N-terminus in the catalytic function of this enzyme; (c) to reveal the structural basis for binding of the zinc finger domain oxidatively-damaged DNA; (d) to elucidate a catalytic mechanism for DNA glycosylate activity; (e) to detect functional groups on Fpg protein and its substrates that facilitate "recognition" of oxidative damage; (f) to establish the structure of complexes formed between Fpg protein or adenine DNA glycosylate and analogs of their DNA substrates; and (g) to study substrate binding and mechanism of action of selected AP endonucleases, and (h) to quantify the contribution of Fpg protein to DNA repair in cells.

这项研究的重点是DNA损伤的生物学后果， 阐明致癌过程中主要事件的总体目标。 一 主要的主题是建立结构之间的关系 受损的DNA和参与DNA复制的酶的功能， 修复. 为此，已经开发了新的实验系统。 位点特异性诱变涉及一种策略，其中穿梭质粒 载体，包含一个单一的定义病变，允许复制， 哺乳动物细胞或细菌。 诱导突变的位置是 通过DNA序列分析确定。 引物延伸反应， 由DNA聚合酶催化，结合稳态动力学分析， 被用来探索translesional合成和诱变事件， 体外 我们的具体目标是（a）建立移码突变模型 在错位的DNA模板和动力学方面， 合成;（B）阐明序列背后的分子基础 背景对碱基替换和缺失的影响;（c）理解 SOS功能在跨病灶合成中的作用;（d）发现 双链修复过程中产生突变的途径 DNA中的无碱基位点;（e）证明DNA之间的差异 聚合酶在其产生由DNA引起的突变的能力方面具有优势， （f）开发预测致突变特异性的体外试验 （g）探索DNA损伤的机制， 损伤增加了哺乳动物中同源重组的频率 细胞和细菌;以及（h）建立以下的溶液结构： 在缺失诱变过程中形成的未对齐中间体。 设计了额外的研究（a）以确定底物 Fpg蛋白的特异性;（B）确定N末端的作用 （c）在该酶的催化功能中;（c）揭示结构 锌指结构域氧化损伤DNA结合的基础;（d） 阐明DNA糖基化活性的催化机制;（e） 检测Fpg蛋白及其底物上的官能团， 促进氧化损伤的“识别”;（f）建立 Fpg蛋白与腺嘌呤DNA形成复合物的结构 糖基化物及其DNA底物的类似物;以及（g）研究 底物结合和所选AP核酸内切酶的作用机制， 和（h）定量Fpg蛋白对DNA修复的贡献， 细胞

项目成果

Mutagenicity of a unique 8-oxoguanine in a human Ha-ras sequence in mammalian cells.

哺乳动物细胞中人类 Ha-ras 序列中独特的 8-氧代鸟嘌呤的致突变性。

• DOI: 10.1093/carcin/16.11.2779
• 发表时间: 1995
• 期刊: Carcinogenesis
• 影响因子: 4.7
• 作者: LePage,F;Margot,A;Grollman,AP;Sarasin,A;Gentil,A
• 通讯作者: Gentil,A

Inhibition of cellular thymidylate synthesis by cytotoxic propenal derivatives of pyrimidine bases and deoxynucleosides.

嘧啶碱基和脱氧核苷的细胞毒性丙烯醛衍生物抑制细胞胸苷酸合成。

• DOI: 10.1016/0006-2952(91)90732-k
• 发表时间: 1991
• 期刊: Biochemical pharmacology
• 影响因子: 5.8
• 作者: Kalman,TI;Marinelli,ER;Xu,B;Reddy,AR;Johnson,F;Grollman,AP
• 通讯作者: Grollman,AP

Miscoding properties of model estrogen-DNA adducts in reactions catalyzed by mammalian and Escherichia coli DNA polymerases.

哺乳动物和大肠杆菌 DNA 聚合酶催化反应中模型雌激素-DNA 加合物的错误编码特性。

• DOI: 10.1021/bi962275q
• 发表时间: 1997
• 期刊: Biochemistry.
• 作者: Shibutani,S;Itoh,S;Yoshizawa,I
• 通讯作者: Yoshizawa,I

Mechanism of mutation on DNA templates containing synthetic abasic sites: study with a double strand vector.

含有合成脱碱基位点的 DNA 模板的突变机制：使用双链载体进行研究。

• DOI: 10.1093/nar/22.10.1897
• 发表时间: 1994
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Takeshita,M;Eisenberg,W
• 通讯作者: Eisenberg,W

Structure of the uncomplexed DNA repair enzyme endonuclease VIII indicates significant interdomain flexibility.

未复杂的DNA修复酶内核酸内切核酸酶VIII的结构表明域间柔韧性明显。

• DOI: 10.1093/nar/gki796
• 发表时间: 2005
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Golan G;Zharkov DO;Feinberg H;Fernandes AS;Zaika EI;Kycia JH;Grollman AP;Shoham G
• 通讯作者: Shoham G