Gene trap mutagenesis-based forward genetic approach reveals that the tumor suppressor OVCA1 is a component of the biosynthetic pathway of diphthamide on elongation factor 2
Gene trap mutagenesis-based forward genetic approach reveals that the tumor suppressor OVCA1 is a component of the biosynthetic pathway of diphthamide on elongation factor 2
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DOI：
10.1074/jbc.m413017200
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发表时间：
2005-03-18
影响因子：
4.8
通讯作者：
Miyagawa, K
中科院分区：
文献类型：
--
作者：
Nobukuni, Y;Kohno, K;Miyagawa, K
OVCA 1是通过从染色体17p13.3（乳腺癌和卵巢癌中染色体畸变的热点）定位克隆而鉴定的肿瘤抑制因子。已经表明，OVCA 1的表达在一些肿瘤中减少，并且它调节细胞增殖、胚胎发育和肿瘤发生。然而，OVCA 1的生物化学功能仍然未知。最近，我们从中国仓鼠卵巢细胞的基因陷阱插入突变体库中分离到一个新的抗白喉毒素和假单胞菌外毒素A的突变体。在这个突变体中，Ovca1基因被破坏的基因陷阱诱变，这种破坏与毒素抗性表型。我们证明了直接的证据表明，肿瘤抑制OVCA 1是一个组成部分的生物合成途径的二苯二甲酰胺的延伸因子2，目标的细菌ADP核糖基化毒素。利用哺乳动物细胞的随机基因陷阱突变体库的功能遗传学方法应该成为一个有用的策略，以确定负责特定表型的基因。
OVCA1 is a tumor suppressor identified by positional cloning from chromosome 17p13.3, a hot spot for chromosomal aberration in breast and ovarian cancers. It has been shown that expression of OVCA1 is reduced in some tumors and that it regulates cell proliferation, embryonic development, and tumorigenesis. However, the biochemical function of OVCA1 has remained unknown. Recently, we isolated a novel mutant resistant to diphtheria toxin and Pseudomonas exotoxin A from the gene trap insertional mutants library of Chinese hamster ovary cells. In this mutant, the Ovca1 gene was disrupted by gene trap mutagenesis, and this disruption well correlated with the toxin-resistant phenotype. We demonstrated direct evidence that the tumor suppressor OVCA1 is a component of the biosynthetic pathway of diphthamide on elongation factor 2, the target of bacterial ADP-ribosylating toxins. A functional genetic approach utilizing the random gene trap mutants library of mammalian cells should become a useful strategy to identify the genes responsible for specific phenotypes.