Complementation assisted positional cloning of the gene for AT variant.

AT 变体基因的互补辅助定位克隆。

• 批准号: 10672136
• 负责人: MATSUURA Shinya
• 金额: $ 1.98万
• 依托单位: Hiroshima University
• 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 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672136/
• 关键词: Nijmegen syndrome; Cancer prone disorder; Ataxia-telangiectasia; Radiosensitivity; Positional cloning; NBS1; Xrs2; Nijmegen breakage症候群; Ataxia-telangiectasia; 染色体不安定症候群; NBSI; 8q21; DNA修復; テロメア; 減数分裂

Nijmegen syndrome is an autosomal recessive disorder characterized by microcephaly, growth retardation, severe combined immunodeficiency, and a high incidence of lymphoid cancers. Cells from NBS patients display chromosome instability, hypersensitivity to ionizing radiation, and an abnormal cell cycle regulation after irradiation. We mapped the. NBS gene at 8q21-24 by functional complementation assays using microcell-mediated chromosome transfer. Only a fragment of chromosome 8q complemented the sensitivity to ionizing radiation in NBS cell lines. We then succeeded in YAC complementation in the NBS cell, and substantially narrowed the NBS critical region to an 800 kb interval. Finally, we identified the NBS1 gene, which is mutated in NBS patients. The 5-bp-deletion, 657-661delACAAA, was found to be founder mutation. The gene is 50 kb long and encodes a protein of 754 amino acids. A weak homology in the N-terminal region was identified with the yeast Xrs2 proteins. Expression of the 2.6 kb transcripts is enhanced in the testis. These results suggested that the NBSM gene might be involved in meiotic recombination.

Nijmegen综合征是一种常染色体隐性障碍，其特征是小头畸形，生长迟钝，严重的联合免疫缺陷和淋巴癌的高发病率。来自NBS患者的细胞表现出染色体不稳定性，对电离辐射的过敏性以及照射后的异常细胞周期调节。我们映射了。使用Microcell介导的染色体转移，通过功能互补测定在8Q21-24处的NBS基因。只有8Q染色体的片段补充了NBS细胞系中电离辐射的敏感性。然后，我们成功地在NBS细胞中取得了YAC的补充，并将NBS临界区域缩小到了800 kb的间隔。最后，我们确定了NBS1基因，该基因在NBS患者中被突变。发现5 bp-Demotion，657-661Delacaaa是创始人突变。该基因长50 kb，编码754个氨基酸的蛋白质。用酵母XRS2蛋白鉴定出N末端区域的弱同源性。在睾丸中增强了2.6 kb转录本的表达。这些结果表明，NBSM基因可能参与减数分裂重组。

项目成果

Hiramoto, T., Nakanishi, T., Sumiyoshi, T., Fukuda, T., Matsuura, S., Tauchi, H., Komatsu, K., Shibasaki, M., Sumii, M., Kajiyama, G., Kamada, N., Miyagawa, K., and Kamiya, K.: "Mutations of a novel human RAD54 homologue, RAD54B, in primary cancer."Oncoge

平本 T.、中西 T.、住吉 T.、福田 T.、松浦 S.、田内 H.、小松 K.、柴崎 M.、纯井 M.、梶山 G.、

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Matsuura,S.,et al.：“奈梅亨断裂综合征基因的位置克隆”Nature Genet.. 19. 179-181 (1998)

Matsuura, K., Balmukhanov, T., Tauchi, H., Weemaes. C., Smeets. D., Chrzanowska, K., Endo, S., Matsuura, S., and Komatsu, K.: "Radiation induction of p53 in cells from Nijmegen breakage syndrome is defective but not similar to ataxia-telangiectasia."Bioch

松浦，K.，巴尔穆哈诺夫，T.，陶奇，H.，威马斯。

Hiramoto,T.,et al.: "Mutation of a novel RAD54 homologue,RAD54B,in primary cancer"Oncogene.. 18. 3422-3426 (1999)

Hiramoto,T.,et al.：“原发性癌症中新型 RAD54 同源物 RAD54B 的突变”Oncogene.. 18. 3422-3426 (1999)

Matsuura, S., Tauchi, H., Nakamura, A., Kondo, N. Sakamoto, S., Endo, S., Smeets. D., Solder, B., Belohradsky, B. H., Kaloustian, V., M., Oshimura, MA Isomura. M., Nakamura, Y., and Komatsu, K.: "Positional cloning of the gene for Nijmegen breakage syndro

松浦，S.，田内，H.，中村，A.，近藤，N.坂本，S.，远藤，S.，斯梅茨。