Study of molecular pathogenesis of Fanconi anemia

范可尼贫血的分子发病机制研究

• 批准号: 14570963
• 负责人: YAMASHITA Takayuki
• 金额: $ 2.24万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570963/
• 关键词: Fanconi anemia; DNA repair; ubiquitin ligase; protein interaction; nuclear transport; reversion; Fanconi貧血; 骨髄不全; 熱ショック蛋白; 再生不良性貧血; 骨髄異形成症候群; 白血病; ゲノム不安定性

Fanconi anemia is an autosomal recessive disorder of hematopoiesis with at least 11 genetically different groups, characterized by cellular hypersensitivity to DNA cross-linkers and chromosome instability. To date, 8 genes have been identified. A multiprotein complex including these gene products, FANCAICJE/F/G/L is required for activation of FANCD2 into a monoubiquitinated form. This active form affects genomic instability in collaboration with BRCA1 and BRCA2IFANCD1. In this project, we studied functions of FANCA mutants with amino acid substitution in its N-terminal region including nuclear localization signal and FANCG-binding sites. Our results indicate that formation of a stable complex including FANCAICIE/FIGIL is not essential for FANCD2 activation. It seems that nuclear import of FANCA/L complex palys a central role in this pathway, and that FANCG negatively regulates nuclear import of FANCA. Furthermore, we identified Hsc7O as a FANCA-binding protein. A dominant-negative form of Hsc7O and 17-AAG, a specific inhibitor of Hsp9O, inhibited nuclear localization of FANCA, suggesting that a chaperone complex including Hsc7O is required for nuclear import of FANCA. We identified FANCA mutations in 27 Japanese FA patients and found novel molecular mechanisms of generation of large deletions in this gene. We found that long-term remission of bone marrow failurewas associated with myeloid lineage-selective expansion of cells with reversion in a patient. This case suggests the clinical usefulness of gene therapy.

范可尼贫血是一种常染色体隐性遗传性造血障碍，至少有11个遗传不同的群体，其特征是细胞对DNA交联剂的超敏反应和染色体不稳定性。迄今为止，已经鉴定出8个基因。包括这些基因产物的多蛋白复合物FANCAICJE/F/G/L是将FANCD 2活化成单泛素化形式所必需的。这种活性形式与BRCA 1和BRCA 2 IFANCD 1合作影响基因组不稳定性。本课题研究了N端氨基酸替换的FANCA突变体的功能，包括核定位信号和FANCG结合位点。我们的研究结果表明，包括FANCAICIE/FIGIL的稳定复合物的形成对于FANCD 2活化不是必需的。FANCA/L复合物的核输入在该通路中起着重要作用，FANCG负调控FANCA的核输入。此外，我们确定Hsc 7 O作为FANCA结合蛋白。Hsc 7 O和Hsp 9 O的特异性抑制剂17-AAG的显性负性形式抑制FANCA的核定位，表明包括Hsc 7 O的伴侣复合物是FANCA核输入所需的。我们在27例日本FA患者中发现了FANCA突变，并发现了该基因产生大缺失的新分子机制。我们发现，骨髓衰竭的长期缓解与患者骨髓细胞谱系选择性扩增和逆转有关。这个病例表明了基因治疗的临床实用性。

项目成果

Oda T, Yamashita T et al.: "ABT1-associated protein (ABTAP), a novel nuclear protein conserved from yeast to mammals, represses transcriptional activation by ABT1."J Cell Biochem. (印刷中). (2004)

Oda T、Yamashita T 等人：“ABT1 相关蛋白 (ABTAP) 是一种从酵母到哺乳动物中保守的新型核蛋白，可抑制 ABT1 的转录激活。”J Cell Biochem（出版中）。

Oda T, Yamashita T et al.: "ABT1-associated protein (ABTAP), a novel nuclear protein conserved from yeast to mammals, repressestranscriptional activation by ABT1."J Cell Biochem. (in press).

Oda T、Yamashita T 等人：“ABT1 相关蛋白 (ABTAP) 是一种从酵母到哺乳动物中保守的新型核蛋白，可抑制 ABT1 的转录激活。”J Cell Biochem。

Hamanoue S, Yamashita T et al.: "Persistent hematopoiesis associated with lineage-selective mosaicism in Fanconi anemia"Blood. 102. 5059 (2003)

Hamanoue S、Yamashita T 等人：“范可尼贫血中与谱系选择性嵌合相关的持续造血”血液。

Yagasaki H, Yamashita T et al.: "Two common founder mutations of the Fanconi anemia group G gene FANCG/XRCC9 in the Japanese population"Hum Mutat. 21. 555 (2003)

Yagasaki H、Yamashita T 等人：“日本人群中范可尼贫血 G 组基因 FANCG/XRCC9 的两个常见创始人突变”Hum Mutat。

Yagasaki H, Yamashita T et al.: "Two common founder mutations of the Fanconi anemia group G gene FANCG/XRCC9 in the Japanese population."Hum Mutat. 21. 555 (2003)

Yagasaki H、Yamashita T 等人：“日本人群中范可尼贫血 G 组基因 FANCG/XRCC9 的两个常见创始人突变。”Hum Mutat。