Complex DNA Damage Induced by High Linear Energy Transfer Alpha-Particles and Protons Triggers a Specific Cellular DNA Damage Response.
Complex DNA Damage Induced by High Linear Energy Transfer Alpha-Particles and Protons Triggers a Specific Cellular DNA Damage Response.
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DOI：
10.1016/j.ijrobp.2017.11.012
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发表时间：
2018-03-01
期刊：
影响因子：
--
通讯作者：
Parsons JL
中科院分区：
文献类型：
--
作者：
Carter RJ;Nickson CM;Thompson JM;Kacperek A;Hill MA;Parsons JL
旨在研究在与组蛋白包装形成染色质的细胞 DNA 中识别和处理电离辐射 (IR) 诱导的复杂 DNA 损伤 (CDD) 的精确机制，其中两个或多个 DNA 损伤非常接近。 HeLa 和口咽鳞状细胞癌（UMSCC74A 和 UMSCC6）细胞接受高线性能量转移 (LET) α 粒子或质子照射，而低 LET 质子和 X 射线则不同。在照射后的不同时间点，通过定量蛋白质印迹分析位点特异性组蛋白翻译后修饰； DNA 损伤和修复通过不同版本的彗星测定进行测量；并使用克隆形成测定法确定细胞存活率。筛选了低和高 LET 辐射（特别是质子辐射）后的位点特异性组蛋白翻译后修饰，旨在识别那些对 CDD 敏感的组蛋白。我们证明，赖氨酸 120 上泛素化的组蛋白 H2B (H2Bub) 是在高 LET α 粒子和质子照射后数小时特异性诱导的，而不是低 LET 质子或 X 射线/γ 辐射。这与 CDD 水平升高有关，从而导致细胞存活率降低。我们进一步发现，H2Bub 的调节受到两种 E3 泛素连接酶 MSL2 和 RNF20/RNF40 复合物的控制，这两种酶的消耗会导致加工缺陷和 CDD 进一步持续存在，并导致辐射后细胞存活率进一步降低。这项研究表明，CDD 的信号传导和修复，特别是由高 LET IR 诱导的 CDD 信号传导和修复，是通过 MSL2 和 RNF20/40 催化的 H2Bub 的特异性诱导来协调的，这是一种对辐射后细胞存活有显着贡献的机制。
To investigate the precise mechanism of recognition and processing of ionizing radiation (IR)-induced complex DNA damage (CDD), where two or more DNA lesions are in close proximity, in cellular DNA which is packaged with histones to form chromatin. HeLa and oropharyngeal squamous cell carcinoma (UMSCC74A and UMSCC6) cells were irradiated with high linear energy transfer (LET) α-particles or protons, versus low-LET protons and X rays. At various time points after irradiation, site-specific histone post-translational modifications were analyzed by quantitative Western blotting; DNA damage and repair were measured by different versions of the comet assay; and cell survival was determined using clonogenic assays. Site-specific histone post-translational modifications after low- and high-LET radiation, particularly proton irradiation, were screened, aiming to identify those responsive to CDD. We demonstrate that histone H2B ubiquitylated on lysine 120 (H2Bub) is specifically induced several hours after irradiation in response to high-LET α-particles and protons but not by low-LET protons or X rays/γ-radiation. This is associated with increased levels of CDD, which contributes to decreased cell survival. We further discovered that modulation of H2Bub is under the control of two E3 ubiquitin ligases, MSL2 and RNF20/RNF40 complex, whose depletion leads to defective processing and further persistence of CDD, and to additional decreased cell survival after irradiation. This study demonstrates that the signaling and repair of CDD, particularly induced by high-LET IR is co-ordinated through the specific induction of H2Bub catalyzed by MSL2 and RNF20/40, a mechanism that contributes significantly to cell survival after irradiation.
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影响因子：
2.9
作者：
Sutherland, BM;Bennett, PV;Laval, J
通讯作者：
Laval, J
影响因子：
1
作者：
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通讯作者：
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DOI：
10.1016/j.ijrobp.2014.05.010
发表时间：
2014-09-01
影响因子：
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作者：
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通讯作者：
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影响因子：
--
作者：
Nickson CM;Moori P;Carter RJ;Rubbi CP;Parsons JL
通讯作者：
Parsons JL
影响因子：
16
作者：
Moyal L;Lerenthal Y;Gana-Weisz M;Mass G;So S;Wang SY;Eppink B;Chung YM;Shalev G;Shema E;Shkedy D;Smorodinsky NI;van Vliet N;Kuster B;Mann M;Ciechanover A;Dahm-Daphi J;Kanaar R;Hu MC;Chen DJ;Oren M;Shiloh Y
通讯作者：
Shiloh Y