Mutation analysis of the human radiosensitivity gene ATM and identification of related genes

人类放射敏感性基因ATM突变分析及相关基因鉴定

• 批准号: 10680512
• 负责人: EJIMA Yosuke
• 金额: $ 1.79万
• 依托单位: Hiroshima Prefectural College of Health Sciences (2000)Hiroshima Prefectual College of Health and Welfare (1999)Kyoto University (1998)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10680512/
• 关键词: A-T; ATM; radiosensitivity; mutation; genetic disease; microsatellite instability; chromosome 11; chromosome 7; ヒト11番染色体; HRAD17; ヒト腫瘍; ATM遺伝子; 日本人

The ATM on chromosome 11q22.3 is the gene responsible for the human radiosensitive recessive disease ataxia-telangiectasia (A-T). Analysis of mutations of ATM and its related genes is a major subject of this study. (1) ATM mutations in Japanese A-T patients belonging to 8 different families were analyzed by two methods (REF : restriction endonuclease fingerprinting, PCR-SSCP). REF method detected 75% of mutations and the others were detected by PCR-SSCP.Mutations that lead to exon skipping or premature protein truncation were predominant. The two mutations 4612del165 and 7883del5 were found to be founder-effect mutations that may be predominant in Japanese ATM mutant alleles. (2) ATM mutations in human tumor cell lines were analyzed by PCR-SSCP to search for possible occurrence of somatic ATM mutations in non-AT patients. Analysis of 25 cell lines revealed 50 sequence alterations in 16 cell lines. None of them was identical to the mutations previously reported in A-T patients. The most … More striking feature was a high frequency of deletions within the intronic mononucleotide tracts in the 5 colon tumor cell lines with microsatellite instability. Some were associated with abnormal transcripts, implying that ATM is a novel mutation target of microsatellite instability where abnormal transcripts are generated indirectly by intronic mutations. (3) Mutations of 6 ATM-related genes (hRAD1, hRAD9, hRAD17, hHUS1, CHK1, CHES1 ) were analyzed in human tumor cell lines. Mutation frequency was very low compared with ATM.One missense polymorphism on hRAD17 was particularly intriguing because this site is highly conserved among species. (4) ATM-homologous sequences were analyzed to search for novel ATM-related genes. Southern blot analysis revealed a number of ATM-hybridizing bands outside chromosome 11. Among them, a 5.5-kb EcoRI fragment on chromosome 7 was analyzed. Sequence analysis of a 5442-bp fragment (p7LA5.5) cloned from chromosome 7-specific genomic library indicated 450-bp region around ATM exon 30 is included. This unprocessed pseudogene is supposed to be generated by a retrotransposition of a part of ATM gene. Further analysis of another ATM-hybridizing bands may lead to the identification of a novel functional gene that shares a motif with ATM. Less

染色体11q22.3上的ATM是人类放射敏感隐性疾病共济失调-毛细血管扩张症（A-T）的基因。分析ATM及其相关基因的突变是本研究的主要内容。(1)采用限制性内切酶指纹图谱（REF：限制性内切酶指纹图谱）和PCR-SSCP两种方法分析日本A-T患者8个不同家族的ATM突变。REF法检测到75%的突变，其余的采用PCR-SSCP检测。导致外显子跳跃或过早的蛋白质截断的突变是主要的。发现两个突变4612del165和7883del5是创始效应突变，可能在日本ATM突变等位基因中占主导地位。(2)利用PCR-SSCP分析人肿瘤细胞系中ATM突变，寻找非at患者体内可能发生的体细胞ATM突变。对25个细胞系进行分析，发现16个细胞系有50个序列改变。这些突变与之前在A-T患者中报道的突变都不相同。5种具有微卫星不稳定性的结肠癌细胞系中，内含子单核苷酸束的缺失频率较高。其中一些与异常转录本有关，这意味着ATM是微卫星不稳定性的一个新的突变靶点，其中异常转录本是由内含子突变间接产生的。(3)分析了6个atm相关基因（hRAD1、hRAD9、hRAD17、hHUS1、CHK1、CHES1）在人肿瘤细胞系中的突变。与ATM相比，突变频率非常低。hRAD17上的一个错义多态性特别有趣，因为该位点在物种中高度保守。(4)分析atm同源序列，寻找新的atm相关基因。Southern blot分析显示，11号染色体外存在多个atm杂交带。其中，对7号染色体上5.5 kb的EcoRI片段进行了分析。从7号染色体特异性基因组文库中克隆的5442 bp片段（p7LA5.5）序列分析表明，该片段位于ATM外显子30附近的450 bp区域。这个未经处理的假基因被认为是由ATM基因的一部分反转录而产生的。进一步分析另一个ATM杂交带可能导致鉴定一个新的功能基因，与ATM共享一个基序。少

项目成果

Ejima Y.: "Aberrant splicing of the ATM gene associated with shortening of the intronic mononucleotide tract in human colon tumor cell lines : a novel mutation target of microsatellite instability"International Journal of Cancer. (印刷中). (2000)

Ejima Y.：“与人类结肠肿瘤细胞系中内含子单核苷酸链缩短相关的 ATM 基因的异常剪接：微卫星不稳定性的新突变目标”国际癌症杂志（2000 年出版）。

江島洋介: "ヒトにおけるチェックポイント制御と癌" 実験医学. 16. 1184-1188 (1998)

Yosuke Ejima：“检查点控制和人类癌症”实验医学 16. 1184-1188 (1998)。

Ejima,Y.: "Aberrant splicing of the ATM gene associated with shortening of the intronic mononucleotide tract in human colon tumor cell lines : a novel mutation traget of microsatellite instability"International Journal of Cancer. 86. 262-268 (2000)

Ejima，Y.：“与人结肠肿瘤细胞系中内含子单核苷酸链缩短相关的 ATM 基因的异常剪接：微卫星不稳定性的新突变目标”国际癌症杂志。

Yosuke Ejima: "A New Development in Cell Cycle Study checkpoint control and cancer (Hiroshi Nojima, ed.)"Yodosha. 173 (1998)

Yosuke Ejima：“细胞周期研究检查点控制和癌症的新进展（Hiroshi Nojima，编辑）”Yodosha。

Ejima,Y.: "Aberrant splieing of the ATM gene associated with shortening of the intronic mononucleotide tract in human colon lumor cell lines : a novel mutation target of microsatellite instability"International Journal of Cancer. 86. 262-268 (2000)

Ejima，Y.：“与人结肠肿瘤细胞系中内含子单核苷酸链缩短相关的 ATM 基因的异常剪接：微卫星不稳定性的新突变目标”国际癌症杂志。