Cloning and analysis of the murine Fanconi anemia group C cDNA.
Cloning and analysis of the murine Fanconi anemia group C cDNA.
复制标题
小鼠范可尼贫血 C 组 cDNA 的克隆和分析。
DOI：
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发表时间：
1993
影响因子：
3.5
通讯作者：
M. Buchwald
中科院分区：
文献类型：
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作者：
Rachel Wevrick;Carol A. Clarke;M. Buchwald
范可尼贫血（Fanconi anemia， FA）是一组以细胞水平为特征的疾病之一，其特点是对DNA损伤剂过敏、染色体不稳定和DNA修复缺陷。FA的临床特征包括全血细胞减少症，常伴有特定的先天性畸形，以及易患白血病。由于血液学表现是预后的关键缺陷，FA是基因替代治疗的候选疾病，小鼠模型系统的开发是这项工作的初始阶段必不可少的。先前，我们通过补充FA细胞对DNA交联剂的固有敏感性，克隆了FA C组中的缺陷基因。我们现在已经克隆了人类FACC cDNA的小鼠同源物。小鼠cDNA （Facc）与人类基因产物具有79%的氨基酸序列相似性。小鼠cDNA在人FA(C)细胞中的表达使细胞药物敏感性恢复到正常水平。因此，尽管存在显著的序列差异，但该蛋白的功能仍然是保守的。对小鼠组织RNA的PCR分析显示，该基因在所有成年组织中均有表达，而原位RNA杂交实验显示，该基因在胎儿发育晚期有组织特异性表达。交叉杂交序列存在于其他哺乳动物，鸡和果蝇的DNA中。这些结果支持了一种假设，即FACC基因产物在细胞抵抗DNA损伤剂的基本保护方面起作用，并且这种功能在进化过程中一直保持不变。
Fanconi anemia (FA) is one of a group of disorders characterized at the cellular level by a combination of hypersensitivity to DNA-damaging agents, chromosomal instability, and defective DNA repair. Clinical features of FA include pancytopenia, often accompanied by specific congenital malformations, and a predisposition to leukemia. Since the hematological manifestations are the critical defect in terms of prognosis, FA is a candidate disease for gene replacement therapy, and the development of a mouse model system is essential for the initial stages of this work. Previously, we have cloned the gene defective in FA group C by complementation of the intrinsic sensitivity of FA cells to DNA cross-linking agents. We have now cloned the murine homologue of the human FACC cDNA. The mouse cDNA (Facc) shares 79% amino acid sequence similarity with the human gene product. The expression of the mouse cDNA in human FA(C) cells restores the cellular drug sensitivity to normal levels. Thus, the function of the protein has been conserved despite the significant sequence divergence. PCR analysis of mouse tissue RNA reveals that the gene is expressed in all adult tissues, while in situ RNA hybridization experiments show tissue specific expression at late stages of fetal development. Cross-hybridizing sequences exist in DNA from other mammals, chicken and Drosophila. These results support the hypothesis that the FACC gene product has a role in a basic aspect of cellular protection against DNA damaging agents and that this function has been conserved during evolution.