Henry Wellcome Lab of Cell Biology, College of Medical, Veterinary and Life Sciences, Institute of Molecular, Cell and Systems Biology, University of Glasgow, Glasgow, UK.
The MRC Protein Phosphorylation and Ubiquitylation Unit, The Sir James Black Centre, College of Life Sciences, University of Dundee, Dundee, UK.
EMBO J. 2019 Mar 15;38(6). doi: 10.15252/embj.2018100024. Epub 2019 Feb 25.
NEDD8 is a ubiquitin-like protein that activates cullin-RING E3 ubiquitin ligases (CRLs). Here, we identify a novel role for NEDD8 in regulating the activity of poly(ADP-ribose) polymerase 1 (PARP-1) in response to oxidative stress. We show that treatment of cells with HO results in the accumulation of NEDD8 chains, likely by directly inhibiting the deneddylase NEDP1. One chain type, an unanchored NEDD8 trimer, specifically bound to the second zinc finger domain of PARP-1 and attenuated its activation. In cells in which is deleted, large amounts of tri-NEDD8 constitutively form, resulting in inhibition of PARP-1 and protection from PARP-1-dependent cell death. Surprisingly, these NEDD8 trimers are additionally acetylated, as shown by mass spectrometry analysis, and their binding to PARP-1 is reduced by the overexpression of histone de-acetylases, which rescues PARP-1 activation. Our data suggest that trimeric, acetylated NEDD8 attenuates PARP-1 activation after oxidative stress, likely to delay the initiation of PARP-1-dependent cell death.
NEDD8 是一种泛素样蛋白，可激活 Cullin-RING E3 泛素连接酶 (CRLs)。在这里，我们发现 NEDD8 在调节聚 (ADP-核糖) 聚合酶 1 (PARP-1) 的活性方面具有新的作用，以响应氧化应激。我们表明，用 HO 处理细胞会导致 NEDD8 链的积累，可能是通过直接抑制去泛素化酶 NEDP1。一种链类型，无锚定的 NEDD8 三聚体，特异性结合到 PARP-1 的第二个锌指结构域，并减弱其激活。在 缺失的细胞中，大量的三-NEDD8 持续形成，导致 PARP-1 抑制和 PARP-1 依赖性细胞死亡的保护。令人惊讶的是，如质谱分析所示，这些 NEDD8 三聚体还被乙酰化，其与 PARP-1 的结合通过组蛋白去乙酰化酶的过表达减少，这挽救了 PARP-1 的激活。我们的数据表明，三聚合体、乙酰化的 NEDD8 在氧化应激后减弱 PARP-1 的激活，可能延迟 PARP-1 依赖性细胞死亡的开始。
