Center for Comparative Oncology, University of California, Davis, California 95616, USA.
Mol Cell Biol. 2010 Feb;30(4):1041-8. doi: 10.1128/MCB.01198-09. Epub 2009 Dec 14.
DNA polymerase eta (PolH), a Y family translesion polymerase, is required for repairing UV-induced DNA damage, and loss of PolH is responsible for early onset of malignant skin cancers in patients with xeroderma pigmentosum variant (XPV), an autosomal recessive disorder. Here, we show that PolH, a target of the p53 tumor suppressor, is a short-half-life protein. We found that PolH is degraded by proteasome, which is enhanced upon UV irradiation. We also found that PolH interacts with Pirh2 E3 ligase, another target of the p53 tumor suppressor, via the polymerase-associated domain in PolH and the RING finger domain in Pirh2. In addition, we show that overexpression of Pirh2 decreases PolH protein stability, whereas knockdown of Pirh2 increases it. Interestingly, we found that PolH is recruited by Pirh2 and degraded by 20S proteasome in a ubiquitin-independent manner. Finally, we observed that Pirh2 knockdown leads to accumulation of PolH and, subsequently, enhances the survival of UV-irradiated cells. We postulate that UV irradiation promotes cancer formation in part by destabilizing PolH via Pirh2-mediated 20S proteasomal degradation.
DNA 聚合酶 eta(PolH)是一种 Y 家族的跨损伤聚合酶,对于修复 UV 诱导的 DNA 损伤至关重要。失活 PolH 会导致色素性干皮病变异型(XPV)患者的恶性皮肤癌的早发性发生,XPV 是一种常染色体隐性遗传疾病。在这里,我们发现 PolH 是 p53 肿瘤抑制因子的靶标,是一种半衰期短的蛋白质。我们发现 PolH 可被蛋白酶体降解,而蛋白酶体在 UV 照射下会增强。我们还发现 PolH 通过 PolH 的聚合酶相关结构域和 Pirh2 的 RING 指结构域与另一个 p53 肿瘤抑制因子的靶标 Pirh2 E3 连接酶相互作用。此外,我们表明过表达 Pirh2 会降低 PolH 蛋白的稳定性,而敲低 Pirh2 则会增加其稳定性。有趣的是,我们发现 PolH 被 Pirh2 募集并以非泛素依赖的方式通过 20S 蛋白酶体降解。最后,我们观察到 Pirh2 敲低会导致 PolH 积累,随后增强了 UV 照射细胞的存活。我们推测,UV 照射通过 Pirh2 介导的 20S 蛋白酶体降解破坏 PolH,从而促进癌症的形成。