Kobayashi Yosuke, Watanabe Miho, Okada Yuki, Sawa Hirofumi, Takai Hiroyuki, Nakanishi Makoto, Kawase Yosuke, Suzuki Hiroshi, Nagashima Kazuo, Ikeda Kyoji, Motoyama Noboru
Department of Geriatric Research, National Institute for Longevity Sciences, Obu, Aichi 474-8522, Japan.
Mol Cell Biol. 2002 Apr;22(8):2769-76. doi: 10.1128/MCB.22.8.2769-2776.2002.
A growing number of DNA polymerases have been identified, although their physiological function and relation to human disease remain mostly unknown. DNA polymerase lambda (Pol lambda; also known as Pol beta2) has recently been identified as a member of the X family of DNA polymerases and shares 32% amino acid sequence identity with DNA Pol beta within the polymerase domain. With the use of homologous recombination, we generated Pol lambda(-/-) mice. Pol lambda(-/-) mice develop hydrocephalus with marked dilation of the lateral ventricles and exhibit a high rate of mortality after birth, although embryonic development appears normal. Pol lambda(-/-) mice also show situs inversus totalis and chronic suppurative sinusitis. The surviving male, but not female, Pol lambda(-/-) mice are sterile as a result of spermatozoal immobility. Microinjection of sperm from male Pol lambda(-/-) mice into oocytes gives rise to normal offspring, suggesting that the meiotic process is not impaired. Ultrastructural analysis reveals that inner dynein arms of cilia from both the ependymal cell layer and respiratory epithelium are defective, which may underlie the pathogenesis of hydrocephalus, situs inversus totalis, chronic sinusitis, and male infertility. Sensitivity of Pol lambda(-/-) cells to various kinds of DNA damage is indistinguishable from that of Pol lambda(+/+) cells. Collectively, Pol lambda(-/-) mice may provide a useful model for clarifying the pathogenesis of immotile cilia syndrome.
尽管DNA聚合酶的生理功能及其与人类疾病的关系大多仍不清楚,但已鉴定出越来越多的DNA聚合酶。DNA聚合酶λ(Pol λ;也称为Pol β2)最近被鉴定为DNA聚合酶X家族的成员,在聚合酶结构域内与DNA Pol β具有32%的氨基酸序列同一性。利用同源重组,我们培育出了Pol λ(-/-)小鼠。Pol λ(-/-)小鼠会出现脑积水,侧脑室明显扩张,出生后死亡率很高,不过胚胎发育似乎正常。Pol λ(-/-)小鼠还表现出全内脏反位和慢性化脓性鼻窦炎。存活的雄性(而非雌性)Pol λ(-/-)小鼠因精子不动而不育。将雄性Pol λ(-/-)小鼠的精子显微注射到卵母细胞中可产生正常后代,这表明减数分裂过程未受损。超微结构分析显示,室管膜细胞层和呼吸道上皮细胞纤毛的内动力臂存在缺陷,这可能是脑积水、全内脏反位、慢性鼻窦炎和男性不育症发病机制的基础。Pol λ(-/-)细胞对各种DNA损伤的敏感性与Pol λ(+/+)细胞没有区别。总的来说,Pol λ(-/-)小鼠可能为阐明不动纤毛综合征的发病机制提供一个有用的模型。