3D-structured illumination microscopy reveals clustered DNA double-strand break formation in widespread γH2AX foci after high LET heavy-ion particle radiation.
3D-structured illumination microscopy reveals clustered DNA double-strand break formation in widespread γH2AX foci after high LET heavy-ion particle radiation.
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DOI：
10.18632/oncotarget.22679
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发表时间：
2017-12-12
期刊：
影响因子：
--
通讯作者：
Shibata A
中科院分区：
文献类型：
--
作者：
Hagiwara Y;Niimi A;Isono M;Yamauchi M;Yasuhara T;Limsirichaikul S;Oike T;Sato H;Held KD;Nakano T;Shibata A
电离辐射诱导的DNA双链断裂（DSB）被认为是遗传毒性突变和细胞死亡的主要原因。虽然DSB在X射线或γ射线照射后分散在整个染色质中，但在高线性能量转移（LET）粒子照射后，在1-2个螺旋DNA转角内可产生多种类型的DNA损伤，包括DSB、单链断裂和碱基损伤，定义为复杂的DNA损伤。除了复杂的DNA损伤的形成，最近的证据表明，多个DSB可以紧密地产生沿着轨道的高LET粒子照射。在本文中，通过使用三维（3D）结构照明显微镜，我们确定了高LET碳离子辐照后形成的三维广泛γ H2 AX焦点。G2期细胞中的大γ H2 AX灶包含多个复制蛋白A（RPA）灶，RPA是同源重组期间发生切除的DSB的标志物。此外，我们通过3D分析证明，γ H2 AX焦点内两个单独RPA焦点之间的距离约为700 nm。总之，我们的研究结果表明，高LET重离子粒子诱导成簇DSB形成的规模约为1 μm3。这些紧密定位的DSB被认为是重离子辐照后形成染色体重排的风险。
DNA double-strand breaks (DSBs) induced by ionising radiation are considered the major cause of genotoxic mutations and cell death. While DSBs are dispersed throughout chromatin after X-rays or γ-irradiation, multiple types of DNA damage including DSBs, single-strand breaks and base damage can be generated within 1–2 helical DNA turns, defined as a complex DNA lesion, after high Linear Energy Transfer (LET) particle irradiation. In addition to the formation of complex DNA lesions, recent evidence suggests that multiple DSBs can be closely generated along the tracks of high LET particle irradiation. Herein, by using three dimensional (3D)-structured illumination microscopy, we identified the formation of 3D widespread γH2AX foci after high LET carbon-ion irradiation. The large γH2AX foci in G2-phase cells encompassed multiple foci of replication protein A (RPA), a marker of DSBs undergoing resection during homologous recombination. Furthermore, we demonstrated by 3D analysis that the distance between two individual RPA foci within γH2AX foci was approximately 700 nm. Together, our findings suggest that high LET heavy-ion particles induce clustered DSB formation on a scale of approximately 1 μm3. These closely localised DSBs are considered to be a risk for the formation of chromosomal rearrangement after heavy-ion irradiation.
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影响因子：
4.7
作者：
Averbeck NB;Topsch J;Scholz M;Kraft-Weyrather W;Durante M;Taucher-Scholz G
通讯作者：
Taucher-Scholz G
影响因子：
4.6
作者：
Oike T;Niimi A;Okonogi N;Murata K;Matsumura A;Noda SE;Kobayashi D;Iwanaga M;Tsuchida K;Kanai T;Ohno T;Shibata A;Nakano T
通讯作者：
Nakano T
影响因子：
4.6
作者：
Amornwichet N;Oike T;Shibata A;Nirodi CS;Ogiwara H;Makino H;Kimura Y;Hirota Y;Isono M;Yoshida Y;Ohno T;Kohno T;Nakano T
通讯作者：
Nakano T
影响因子：
4.6
作者：
Kobayashi D;Oike T;Shibata A;Niimi A;Kubota Y;Sakai M;Amornwhichet N;Yoshimoto Y;Hagiwara Y;Kimura Y;Hirota Y;Sato H;Isono M;Yoshida Y;Kohno T;Ohno T;Nakano T
通讯作者：
Nakano T
影响因子：
78.5
作者：
Pageau, Gayle J.;Hall, Lisa L.;Lawrence, Jeanne B.
通讯作者：
Lawrence, Jeanne B.