Breakthrough for a DNA break-preventer.

Breakthrough for a DNA break-preventer.
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DNA 断裂预防剂的突破。

DOI:
10.1073/pnas.1400512111
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发表时间:
2014
影响因子:
11.1
通讯作者:
Lange,SabineS
Lange,SabineS
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Wood,RichardD;Lange,SabineS

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哺乳动物基因组编码大约16种不同的DNA聚合酶,参与DNA复制、DNA修复、重组或绕过DNA损伤的不同方面(1,2)。用于正常复制的DNA聚合酶通常不能在受损的DNA上进行。当遇到受损部位时,复制至少暂时停止,通过调用由专门的DNA聚合酶介导的跨病变DNA合成(TLS)过程,或切换到另一个未受损的DNA模板,可以绕过损伤。在《美国国家科学院院刊》(3)上的一篇报告中,Lee等人。通过他们对DNA聚合酶ζ(POLζ)的研究,极大地促进了理解这一过程的可能性,POL DNA聚合酶可以说是专业DNA聚合酶中最重要的。作者报道了一种由四个亚基组成的人POLζ全酶活性形式的纯化,为这一重要酶的详细生化和结构研究提供了可能。来自酿酒酵母的Polζ已被发现,但哺乳动物的蛋白质具有额外的功能(例如,它在哺乳动物中是必需的,但在酵母中不是),其催化亚基(称为REV3L)大约是酵母蛋白的两倍大小。Polζ在保护哺乳动物细胞免受DNA损伤方面具有特别重要的作用。POLζ是通过TLS绕过许多dna损伤所必需的,尽管如果在dna模板中错误的碱基对面插入错误的碱基可能会发生突变(4-6)。然而,延迟旁路的一个危险后果是,DNA复制叉处的机械可能会崩溃,使其暴露在细胞中的酶下,这些酶切割DNA并形成双链断裂。POLζ有助于复制一些天生难以穿越的自然产生的脱氧核糖核酸序列,例如哺乳动物基因组中的“脆弱部位”区域,或形成非BDNA结构的序列(7,8)。
Mammalian genomes encode about 16 distinct DNA polymerases that participate in different aspects of DNA replication, DNA repair, recombination, or bypass of DNA damage (1, 2). The DNA polymerases used for normal replication generally cannot proceed on damaged DNA. When a damaged site is encountered, replication is stalled at least temporarily, and the lesion may be bypassed by invoking a process of translesion DNA synthesis (TLS) mediated by specialized DNA polymerases, or by switching to another undamaged DNA template. In a report in PNAS (3), Lee et al. significantly advance the possibilities for understanding this process by their work on DNA polymerase ζ (Pol ζ), arguably the most important of the specialized DNA polymerases. The authors describe the purification of an active form of the human Pol ζ holoenzyme composed of four subunits, which opens up the possibility for detailed biochemical and structural studies of this essential enzyme. Pol ζ from the yeast Saccharomyces cerevisiae has been available, but the mammalian protein has additional functions (for example, it is essential in mammals, but not in yeast), and the catalytic subunit (called REV3L) is about twice the size of the yeast protein.Pol ζ is exceptionally significant in the defense of mammalian cells against DNA damage. Pol ζ is needed for the bypass of many DNA lesions by TLS, although it can be mutagenic if an incorrect base is inserted opposite a mis-instructional lesion in the DNA template (4–6). However, a perilous consequence of delaying bypass is that the machinery at the DNA replication fork can collapse, leaving it exposed to enzymes in the cell that cut the DNA and form a double-strand break. Pol ζ aids in replication of some naturally occurring DNA sequences that are inherently difficult to traverse, such as the “fragile-site” regions in mammalian genomes, or sequences forming non-B DNA structures (7, 8).
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