A critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage
A critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage
复制标题
DOI：
10.1016/s0960-9822(00)00610-2
复制
发表时间：
2000-07-27
期刊：
影响因子：
9.2
通讯作者：
Bonner, WM
中科院分区：
文献类型：
--
作者：
Paull, TT;Rogakou, EP;Bonner, WM
背景：真核细胞对基因组DNA双链断裂的反应包括将许多因子封存到核灶中。最近，据报道，组蛋白H2 A家族的成员，H2 AX，成为广泛的磷酸化1-3分钟内的DNA损伤和形式focusatbreak sites.Results：在这项工作中，我们研究的作用H2 AX磷酸化的焦点形成的几个修复相关的复合物，并调查可能参与启动这种反应的因素。使用两种不同的方法在人类细胞中产生DNA双链断裂，我们发现修复因子Rad 50和Rad 51在DNA损伤后各自与磷酸化H2 AX（γ-H2 AX）灶共定位。肿瘤抑制基因BRCA 1的产物也与γ-H2 AX共定位，并在Rad 50或Rad 51之前被募集到这些位点。暴露的细胞真菌抑制剂渥曼青霉素消除病灶形成的所有修复因子检查，表明磷酸肌醇（PI）-3家族的蛋白激酶在介导这种反应的作用。Wortmannin治疗只有在足够早地加入以防止γ-H2 AX形成时才有效，这表明γ-H2 AX对于将其他因子募集到DNA损伤位点是必需的。DNA修复缺陷的细胞表现出显着降低的能力，以增加磷酸化的H2 AX在电离辐射的响应，与γ-H2 AX在DNA repair.Conclusions的作用一致：γ-H2 AX傻瓜的模式，这是建立在几分钟内的DNA损伤占模式的Rad 50，Rad 51，和Brca 1灶看到很久以后在恢复过程中的损害。所提出的证据强烈支持γ-H2 AX和PI-3蛋白激酶家族在双链断裂位点的焦点形成中的作用，并表明伴随双链断裂修复的染色质结构变化的可能性。(C)2000 Elsevier Science Ltd，保留所有权利。
Background: The response of eukaryotic cells to double-strand breaks in genomic DNA includes the sequestration of many factors into nuclear foci. Recently it has been reported that a member of the histone H2A family, H2AX, becomes extensively phosphorylated within 1-3 minutes of DNA damage and forms foci at break sites.Results: In this work, we examine the role of H2AX phosphorylation in focus formation by several repair-related complexes, and investigate what factors may be involved in initiating this response. Using two different methods to create DNA double-strand breaks in human cells, we found that the repair factors Rad50 and Rad51 each colocalized with phosphorylated H2AX (gamma-H2AX) foci after DNA damage. The product of the tumor suppressor gene BRCA1 also colocalized with gamma-H2AX and was recruited to these sites before Rad50 or Rad51. Exposure of cells to the fungal inhibitor wortmannin eliminated focus formation by all repair factors examined, suggesting a role for the phosphoinositide (PI)-3 family of protein kinases in mediating this response. Wortmannin treatment was effective only when it was added early enough to prevent gamma-H2AX formation, indicating that gamma-H2AX is necessary for the recruitment of other factors to the sites of DNA damage. DNA repair-deficient cells exhibit a substantially reduced ability to increase the phosphorylation of H2AX in response to ionizing radiation, consistent with a role for gamma-H2AX in DNA repair.Conclusions: The pattern of gamma-H2AX fool that is established within a few minutes of DNA damage accounts for the patterns of Rad50, Rad51, and Brca1 foci seen much later during recovery from damage. The evidence presented strongly supports a role for the gamma-H2AX and the PI-3 protein kinase family in focus formation at sites of double-strand breaks and suggests the possibility of a change in chromatin structure accompanying double-strand break repair. (C) 2000 Elsevier Science Ltd, All rights reserved.