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Probing ATR activation with model DNA templates.

机译:用模型DNA模板探测ATR激活。

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The ATR kinase is a critical upstream component of a checkpoint pathway that responds to many forms of damaged and incompletely replicated DNA. Cellular processes such as DNA replication and repair are thought to convert these DNA lesions into a common DNA intermediate that activates this signaling pathway. Indeed, numerous studies have shown that two DNA structures formed during these processes--single-stranded DNA (ssDNA) and junctions between double-stranded DNA (dsDNA) and ssDNA--are important components of the ATR-activating structure. However, an unanswered question is whether primed ssDNA is sufficient for activation of the ATR response. We recently demonstrated that primed ssDNA is sufficient to induce a bona fide checkpoint response in Xenopus egg extracts. This is the first well-defined DNA structure capable of eliciting ATR activation. Using this structure, we examined the contribution of ds/ssDNA junctions and ssDNA to checkpoint activation. Our results indicate the context in which the checkpoint-activating structure is generated may contribute significantly to its signaling properties. Here we discuss the implications of our findings, in the context of other recent work in the field, on our understanding of checkpoint signaling.
机译:ATR激酶是检查点途径的关键上游成分,可对多种形式的受损和复制不完全的DNA作出反应。 DNA复制和修复等细胞过程被认为可以将这些DNA损伤转化为激活该信号通路的常见DNA中间体。实际上,大量研究表明,在这些过程中形成的两个DNA结构-单链DNA(ssDNA)以及双链DNA(dsDNA)和ssDNA之间的连接点-是ATR激活结构的重要组成部分。然而,一个未解决的问题是引发的ssDNA是否足以激活ATR反应。我们最近证明,引发的ssDNA足以在非洲爪蟾卵提取物中诱导真正的检查点反应。这是第一个能够引发ATR激活的明确定义的DNA结构。使用这种结构,我们检查了ds / ssDNA连接和ssDNA对检查点激活的贡献。我们的结果表明生成检查点激活结构的上下文可能对其信号传递特性有重大贡献。在此,我们将在本领域其他近期工作的背景下,讨论我们的发现对理解检查点信号的意义。

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