NUCLEAR FOCI OF MAMMALIAN RAD51 RECOMBINATION PROTEIN IN SOMATIC-CELLS AFTER DNA-DAMAGE AND ITS LOCALIZATION IN SYNAPTONEMAL COMPLEXES
NUCLEAR FOCI OF MAMMALIAN RAD51 RECOMBINATION PROTEIN IN SOMATIC-CELLS AFTER DNA-DAMAGE AND ITS LOCALIZATION IN SYNAPTONEMAL COMPLEXES
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DOI：
10.1073/pnas.92.6.2298
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发表时间：
1995-03-14
影响因子：
11.1
通讯作者：
WARD, DC
中科院分区：
文献类型：
--
作者：
HAAF, T;GOLUB, EI;WARD, DC
酿酒酵母的Rad 51蛋白是大肠杆菌重组酶RecA的结构同源物。在酵母中，Rad 51蛋白是有丝分裂和减数分裂重组以及DNA双链断裂修复所必需的。我们已经使用了针对在大肠杆菌中表达的同源人蛋白HsRad 51的抗体。大肠杆菌，可视化的蛋白质在哺乳动物体细胞和减数分裂细胞的空间分布。在培养的人类细胞中，HsRad 51蛋白集中在核质中的多个离散灶中;它在细胞质和核仁中基本上不存在。用甲磺酸甲酯、紫外线照射或Cs-137照射处理细胞后，具有HsRad 51蛋白免疫荧光的细胞百分比增加;相同细胞显示程序外DNA合成。Rad 51焦点的诱导被转录抑制剂阻断。在小鼠粗线期精母细胞中，Rad 51蛋白的小鼠同源物高度富集在联会复合体中，联会复合体在减数分裂前期期间在配对的同源染色体之间形成。我们的结论是哺乳动物蛋白同源酵母Rad 51参与DNA损伤的修复和减数分裂过程中的重组修复。
Rad51 protein of Saccharomyces cerevisiae is a structural homolog of the Escherichia coli recombination enzyme RecA. In yeast, the Rad51 protein is required for mitotic and meiotic recombination and for repair of double-strand breaks in DNA. We have used antibodies raised against the homologous human protein, HsRad51, expressed in E. coli, to visualize the spatial distribution of the protein in mammalian somatic and meiotic cells. In cultured human cells, the HsRad51 protein is concentrated in multiple discrete foci in the nucleoplasm; it is largely absent from cytoplasm and nucleoli. After treatment of cells With methyl methanesulfonate, ultraviolet irradiation, or Cs-137 irradiation, the percentage of cells with HsRad51 protein immunofluorescence increases; the same cells show unscheduled DNA synthesis. Induction of Rad51 foci is blocked by inhibitors of transcription. In mouse pachytene spermatocytes, the mouse homolog of Rad51 protein is highly enriched in synaptonemal complexes that are formed between the paired homologous chromosomes during meiotic prophase. We conclude that the mammalian proteins homologous to yeast Rad51 are involved in repair of DNA damage and recombinational repair during meiosis.