CLONING & CHARACTERIZING THE XP VARIANT MUTATOR GENE(S)

克隆

• 批准号: 2856321
• 负责人: VERONICA M MAHER
• 金额: $ 19.22万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-01-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2856321
• 关键词: DNA damage; DNA repair; DNA replication; caffeine; complementary DNA; gene induction /repression; molecular cloning; nucleic acid sequence; point mutation; polymerase chain reaction; tissue /cell culture; ultraviolet radiation; xeroderma pigmentosum

DESCRIPTION: Cells from individuals with the disease Xeroderma pigmentosum (XP) are defective in different facets of nucleotide excision repair (NER) with the exception of XP-variant (XP-V) individuals, which appear to have normal NER. The most commonly accepted hypothesis for XP-V is that it is due to a defect in a polymerase-related gene that leads to error-prone bypass of lesions. The principal investigator wishes to clone the XP-V gene in order to better understand the molecular basis of this defect. Evidence is presented bearing on the mechanism(s) responsible for the genetic predisposition to sunlight-induced skin cancer found in xeroderma pigmentosum variant (XP-V) patients, who develop clinical features of classical XP patients yet have normal excision repair. Their cells are abnormally slow in replicating UV-damaged DNA and are extremely sensitive to UV-induced mutations. Data has been obtained that suggest that this extreme hypermutability reflects a defective, error-prone replication complex that yields an abnormal spectrum of UV-induced mutations. Not only is replication in the intact cells abnormally error-prone but this is also true for in vitro replication of an irradiated template by cell-free extracts from XP-V cells. They represent the only known example of human cells with a defective error-prone replication complex not resulting from defective DNA repair. Thus, they offer an opportunity to investigate the mechanisms normal cells use to insure high-fidelity DNA replication. It is proposed to use two approaches simultaneously to clone the gene(s) responsible for the abnormal characteristics of XP-V cells and then to characterize the gene identified and its protein product.

描述：细胞来自患有着色性干皮病的个体 (XP)在核苷酸切除修复（NER）的不同方面有缺陷 除了XP变异（XP-V）个体外， 正常NER。 对于XP-V最普遍接受的假设是， 由于聚合酶相关基因的缺陷， 病变旁路。 首席研究员希望克隆XP-V基因 以便更好地理解这种缺陷的分子基础。 证据 与遗传学的机制有关， 在干皮病中发现的日光诱发皮肤癌的易感性 色素变性（XP-V）患者，这些患者出现以下临床特征 经典XP患者仍具有正常的切除修复。 他们的细胞 在复制紫外线损伤的DNA时异常缓慢，并且对 紫外线诱导的突变 数据显示，这种极端 超变性反映了一种有缺陷的、容易出错的复制复合体， 产生了异常的紫外线诱导突变谱。 不仅 在完整的细胞中复制异常容易出错，但这也是真的 用无细胞提取物体外复制辐照过的模板 来自XP-V细胞。 它们代表了唯一已知的人类细胞的例子， 不是由有缺陷的DNA引起的有缺陷的易错复制复合体 修复. 因此，他们提供了一个机会，调查机制， 正常细胞用来确保高保真的DNA复制。 提出要 同时使用两种方法来克隆负责 XP-V细胞的异常特征，然后表征该基因 鉴定及其蛋白质产物。

项目成果

Abnormal, error-prone bypass of photoproducts by xeroderma pigmentosum variant cell extracts results in extreme strand bias for the kinds of mutations induced by UV light.

着色性干皮病变异细胞提取物对光产物的异常且容易出错的旁路导致了紫外线诱导的突变类型的极端链偏差。

• DOI: 10.1128/mcb.19.1.147
• 发表时间: 1999
• 期刊: Molecular and cellular biology
• 影响因子: 5.3
• 作者: McGregor,WG;Wei,D;Maher,VM;McCormick,JJ
• 通讯作者: McCormick,JJ

Relationship between adduct formation, rates of excision repair and the cytotoxic and mutagenic effects of structurally-related polycyclic aromatic carcinogens.

加合物形成、切除修复率与结构相关多环芳香族致癌物的细胞毒性和致突变作用之间的关系。

• DOI: 10.1016/s0027-5107(97)00037-7
• 发表时间: 1997
• 期刊: Mutation research
• 作者: McGregor,WG;Wei,D;Chen,RH;Maher,VM;McCormick,JJ
• 通讯作者: McCormick,JJ

Effect of nuclear environment on the distribution of benzo[a]pyrene diol epoxide-induced adducts in the HPRT gene of human fibroblasts.

核环境对苯并[a]芘二醇环氧化物诱导的加合物在人成纤维细胞HPRT基因中分布的影响。

• DOI: 10.1093/carcin/17.12.2695
• 发表时间: 1996
• 期刊: Carcinogenesis
• 影响因子: 4.7
• 作者: Wei,D;Maher,VM;McCormick,JJ
• 通讯作者: McCormick,JJ

Site-specific excision repair of 1-nitrosopyrene-induced DNA adducts at the nucleotide level in the HPRT gene of human fibroblasts: effect of adduct conformation on the pattern of site-specific repair.

人成纤维细胞 HPRT 基因核苷酸水平上 1-亚硝基芘诱导的 DNA 加合物的位点特异性切除修复：加合物构象对位点特异性修复模式的影响。

• DOI: 10.1128/mcb.16.7.3714
• 发表时间: 1996
• 期刊: Molecular and cellular biology
• 影响因子: 5.3
• 作者: Wei,D;Maher,VM;McCormick,JJ
• 通讯作者: McCormick,JJ

Site-specific rates of excision repair of benzo[a]pyrene diol epoxide adducts in the hypoxanthine phosphoribosyltransferase gene of human fibroblasts: correlation with mutation spectra.

• DOI: 10.1073/pnas.92.6.2204
• 发表时间: 1995-03
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: D. Wei;V. Maher;J. Mccormick