Characterization of a Y-Family DNA Polymerase eta from the Eukaryotic Thermophile Alvinella pompejana
Characterization of a Y-Family DNA Polymerase eta from the Eukaryotic Thermophile Alvinella pompejana
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DOI：
10.4061/2010/701472
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发表时间：
2010-01-01
影响因子：
2.3
通讯作者：
Iwai, Shigenori
中科院分区：
文献类型：
--
作者：
Kashiwagi, Sayo;Kuraoka, Isao;Iwai, Shigenori
人DNA聚合酶eta（HsPol.）在跨损伤合成（TLS）中起重要作用，TLS允许复制过去的DNA损伤，如UV诱导的顺式-顺式环丁烷嘧啶二聚体（CPD）。在这里，我们的特点ApPol eta从嗜热蠕虫Alvinella pompejana，居住在深海热液喷口烟囱。ApPol eta与HsPol eta具有序列同源性，并含有结合泛素和增殖细胞核抗原的结构域。太阳诱导的紫外线不会穿透Alvinella的环境;然而，这种新型DNA聚合酶催化了有效和准确的TLS通过CPD，以及活性氧诱导的7，8-二氢-8-氧代鸟嘌呤和胸腺嘧啶二醇的异构体。此外，我们发现ApPol eta比HsPol eta更耐热，正如其栖息地温度所预期的那样。此外，该酶的活性在较高浓度的有机溶剂存在下得以保留。因此，ApPol eta提供了一种稳健的、类似人类的Pol eta，在暴露于高温和有机溶剂后更具活性。
Human DNA polymerase eta (HsPol.) plays an important role in translesion synthesis (TLS), which allows for replication past DNA damage such as UV-induced cis-syn cyclobutane pyrimidine dimers (CPDs). Here, we characterized ApPol eta from the thermophilic worm Alvinella pompejana, which inhabits deep-sea hydrothermal vent chimneys. ApPol eta shares sequence homology with HsPol eta and contains domains for binding ubiquitin and proliferating cell nuclear antigen. Sun-induced UV does not penetrate Alvinella's environment; however, this novel DNA polymerase catalyzed efficient and accurate TLS past CPD, as well as 7,8-dihydro-8oxoguanine and isomers of thymine glycol induced by reactive oxygen species. In addition, we found that ApPol eta is more thermostable than HsPol eta, as expected from its habitat temperature. Moreover, the activity of this enzyme was retained in the presence of a higher concentration of organic solvents. Therefore, ApPol eta provides a robust, human-like Pol eta that is more active after exposure to high temperatures and organic solvents.