Proximity RNA labeling reveals functions of lncRNA in DNA damage response
Proximity RNA labeling reveals functions of lncRNA in DNA damage response
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DOI：
10.1007/s42764-023-00099-0
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发表时间：
2023-05
期刊：
影响因子：
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通讯作者：
Weili Li;Zhongxia Li;Zhiwen Deng;J. Zhai;Shuzhen Han;Xiangyu Liu
中科院分区：
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作者：
Weili Li;Zhongxia Li;Zhiwen Deng;J. Zhai;Shuzhen Han;Xiangyu Liu
长非编码RNA（lncRNA）正在成为保持基因组稳定性并与包括癌症在内的人类疾病相关的重要参与者（Statello et al.，Statello等人，Nature Reviews Molecular Cell Biology 22：96 - 118，2021）。然而，lncRNA促进DNA损伤反应的潜在机制尚不完全清楚。其中一个局限性是，目前的研究依赖于lncRNA和诱饵之间的物理相互作用，因此可能无法检测到弱相互作用。在这里，我们应用了一种方法来通过抗体介导的蛋白A-抗坏血酸过氧化物酶2（APEX2）标记gH2ax（DNA损伤的标志物）的近端lncRNA，并发现几种lncRNA共定位在DNA损伤位点内，包括结合BARD 1的BGL 3和与DNA-PK相互作用的SNHG 12（Haemmig等人，Haemmig等人，Sci Transl Med，2020; Hu等人，Hu等人，EMBO Journal 39：e104133，2020）。此外，我们证明了SNHG12的敲低对人宫颈癌HeLa和结肠癌HCT 116细胞的辐射敏感性。总之，我们的研究为探讨lncRNA在DNA损伤反应中的作用提供了一种新的方法。
Long non-coding RNAs (lncRNA) are emerging as important players to keep genome stability and associate with human diseases including cancers (Statello et al., Statello et al., Nature Reviews Molecular Cell Biology 22:96–118, 2021). However, the underlying mechanisms that lncRNAs contribute to DNA damage response are not fully known. One of the limitations is that current studies depend on the physical interaction between lncRNA and the decoy, so that weak interactions might not be detected. Here we applied a method to label proximal lncRNAs of gH2ax, a marker of DNA damage, by antibody-mediated protein A-ascorbate peroxidase 2 (APEX2) and discovered that several lncRNA co-localized within DNA damage sites, including BGL3 which binds BARD1 and SNHG12 which interacts with DNA-PK (Haemmig et al., Haemmig et al., Sci Transl Med, 2020; Hu et al., Hu et al., EMBO Journal 39:e104133, 2020). In addition, we proved that knockdown of SNHG12 sensitized human cervical cancer HeLa and colon cancer HCT116 cells to irradiation. Overall, our study provides a new method to explore the function of lncRNA in DNA damage response.