Department of Microbiology, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, SP, Brazil.
NUPEB & Biological Sciences Department, Federal University of Ouro Preto, Ouro Preto, MG, Brazil.
Photochem Photobiol. 2019 Jan;95(1):345-354. doi: 10.1111/php.13041. Epub 2018 Nov 23.
The crucial role of DNA polymerase eta in protecting against sunlight-induced tumors is evidenced in Xeroderma Pigmentosum Variant (XP-V) patients, who carry mutations in this protein and present increased frequency of skin cancer. XP-V cellular phenotypes may be aggravated if proteins of DNA damage response (DDR) pathway are blocked, as widely demonstrated by experiments with UVC light and caffeine. However, little is known about the participation of DDR in XP-V cells exposed to UVA light, the wavelengths patients are mostly exposed. Here, we demonstrate the participation of ATR kinase in protecting XP-V cells after receiving low UVA doses using a specific inhibitor, with a remarkable increase in sensitivity and γH2AX signaling. Corroborating ATR participation in UVA-DDR, a significant increase in Chk1 protein phosphorylation, as well as S-phase cell cycle arrest, is also observed. Moreover, the participation of oxidative stress is supported by the antioxidant action of N-acetylcysteine (NAC), which significantly protects XP-V cells from UVA light, even in the presence of the ATR inhibitor. These findings indicate that the ATR/Chk1 pathway is activated to control UVA-induced oxidatively generated DNA damage and emphasizes the role of ATR kinase as a mediator of genomic stability in pol eta defective cells.
在 Xeroderma Pigmentosum Variant(XP-V)患者中,DNA 聚合酶 eta 在预防阳光诱导的肿瘤中起着至关重要的作用,这些患者携带该蛋白的突变,皮肤癌的发病率增加。如果 DNA 损伤反应(DDR)途径的蛋白质被阻断,如广泛的 UVC 光和咖啡因实验所证明的那样,XP-V 细胞表型可能会加重。然而,对于 XP-V 细胞在接受低剂量 UVA 光照射时 DDR 的参与,人们知之甚少,而这些波长是患者最容易暴露的。在这里,我们使用特异性抑制剂证明了 ATR 激酶在接受低剂量 UVA 照射后保护 XP-V 细胞的参与,同时显著增加了敏感性和 γH2AX 信号。ATR 参与 UVA-DDR 的佐证是 Chk1 蛋白磷酸化的显著增加,以及 S 期细胞周期停滞。此外,抗氧化应激的参与得到了 N-乙酰半胱氨酸(NAC)的抗氧化作用的支持,即使在 ATR 抑制剂存在的情况下,NAC 也能显著保护 XP-V 细胞免受 UVA 光的伤害。这些发现表明,ATR/Chk1 途径被激活以控制 UVA 诱导的氧化性 DNA 损伤,并强调了 ATR 激酶作为 pol eta 缺陷细胞中基因组稳定性的介导者的作用。