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BRCTx Is a Novel, Highly Conserved RAD18-Interacting Protein

机译:BRCTx是一种新型的高度保守的RAD18相互作用蛋白。

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The BRCT domain is a highly conserved module found in many proteins that participate in DNA damage checkpoint regulation, DNA repair, and cell cycle control. Here we describe the cloning, characterization, and targeted mutagenesis of Brctx, a novel gene with a BRCT motif. Brctx was found to be expressed ubiquitously in adult tissues and during development, with the highest levels found in testis. Brctx-deficient mice develop normally, show no pathological abnormalities, and are fertile. BRCTx binds to the C terminus of hRAD18 in yeast two-hybrid and immunoprecipitation assays and colocalizes with this protein in the nucleus. Despite this, Brctx-deficient murine embryonic fibroblasts (MEFs) do not show overt sensitivity to DNA-damaging agents. MEFs from Brctx-deficient embryos grow at a similar rate to wild-type MEF CD4/CD8 expressions, and the cell cycle parameters of thymocytes from wild-type and Brctx knockout animals are indistinguishable. Intriguingly, the BRCT domain of BRCTx is responsible for mediating its localization to the nucleus and centrosome in interphase cells. We conclude that, although highly conserved, Brctx is not essential for the above-mentioned processes and may be redundant.
机译:BRCT结构域是在许多蛋白质中发现的高度保守的模块,这些蛋白质参与DNA损伤检查点调节,DNA修复和细胞周期控制。在这里,我们描述了 Brctx (一种具有BRCT主题的新型基因)的克隆,表征和定向诱变。发现 Brctx 在成人组织和发育过程中普遍表达,在睾丸中的表达最高。 Brctx 缺陷小鼠正常发育,无病理异常,可育。在酵母双杂交和免疫沉淀试验中,BRCTx与hRAD18的C末端结合,并与该蛋白质在细胞核中共定位。尽管如此, Brctx 缺陷型鼠胚胎成纤维细胞(MEF)对DNA破坏剂没有明显的敏感性。来自 Brctx 缺陷胚胎的MEF以与野生型MEF CD4 / CD8表达相似的速率生长,并且野生型和 Brctx 敲除动物胸腺细胞的细胞周期参数难以区分。有趣的是,BRCTx的BRCT结构域负责介导其在相间细胞中的核和中心体的定位。我们得出的结论是,尽管 Brctx 高度保守,但对于上述过程并不是必需的,并且可能是多余的。

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