Department of Genetics, Erasmus Medical Centre, Rotterdam 3015 GE, The Netherlands.
Mol Cell. 2013 Aug 22;51(4):469-79. doi: 10.1016/j.molcel.2013.08.007.
Chromatin remodeling is tightly linked to all DNA-transacting activities. To study chromatin remodeling during DNA repair, we established quantitative fluorescence imaging methods to measure the exchange of histones in chromatin in living cells. We show that particularly H2A and H2B are evicted and replaced at an accelerated pace at sites of UV-induced DNA damage. This accelerated exchange of H2A/H2B is facilitated by SPT16, one of the two subunits of the histone chaperone FACT (facilitates chromatin transcription) but largely independent of its partner SSRP1. Interestingly, SPT16 is targeted to sites of UV light-induced DNA damage-arrested transcription and is required for efficient restart of RNA synthesis upon damage removal. Together, our data uncover an important role for chromatin dynamics at the crossroads of transcription and the UV-induced DNA damage response.
染色质重塑与所有 DNA 转导活性密切相关。为了研究 DNA 修复过程中的染色质重塑，我们建立了定量荧光成像方法来测量活细胞中染色质组蛋白的交换。我们发现，特别是在 UV 诱导的 DNA 损伤部位，H2A 和 H2B 被迅速逐出并取代。这种 H2A/H2B 的快速交换是由组蛋白伴侣 FACT（促进染色质转录）的两个亚基之一 SPT16 介导的，但在很大程度上独立于其伴侣 SSRP1。有趣的是，SPT16 被靶向到 UV 光诱导的 DNA 损伤停滞转录的部位，并且在去除损伤后重新启动 RNA 合成是必需的。总之，我们的数据揭示了染色质动力学在转录和 UV 诱导的 DNA 损伤反应交叉点的重要作用。
