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菌根Ku和酶蛋白构成一个两部分的NHEJ修复机器
Marina Della1, Phillip L Palmbos, Hui-Min Tseng
1Cambridge Institute for Medical Research, University of Cambridge, Department of Haematology, Hills Road, Cambridge CB2 2XY, UK.
Science (New York, N.Y.)
|October 23, 2004
概括
Prokaryotic DNA 连接酶 D (Mt-Lig) 和 Ku 蛋白质可以使用非同类末端连接 (NHEJ) 修复 DNA 双链断裂 (DSB). 这种细菌系统对基因组稳定性进行关键的终端处理和结合.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 微生物学 微生物学
背景情况:
- 非同类末端结合 (NHEJ) 对哺乳动物基因组稳定至关重要,它可以修复DNA双链断裂 (DSB).
- 虽然NHEJ结合已被理解,但之前的最终处理步骤在很大程度上仍然没有特征.
- 最近,在 prokaryotes 中发现了同源的末端桥梁和结活动.
研究的目的:
- 为了研究来自Mycobacterium tuberculosis (Mt-Lig) 的DNA结合酶D的酶活性.
- 为了确定 prokaryotic 蛋白质是否可以重组非同类末端连接 (NHEJ) 以进行 DNA 双链断裂 (DSB) 修复.
主要方法:
- 生物化学试验被用来描述Mt-Lig.的核样转移酶和外核酶活动.
- 在体外实验中将Mt-Ku和Mt-Lig结合起来,以评估它们连接不兼容的DSB端的能力.
- 在酵母体内的体内实验测试了Mt-Ku和Mt-Lig重构NHEJ的能力.
主要成果:
- Mt-Lig表现出不同的核基转移酶活动 (聚合酶,终端转移酶,原酶) 和3'至5'外核酶活动.
- Mt-Ku和Mt-Lig的组合成功连接不兼容的DSB在体外结束.
- 这些 prokaryotic 蛋白质在酵母细胞中重建了功能性的 NHEJ.
结论:
- Prokaryotic Ku 和 DNA 连接酶 D 构成一个完整的非同类末端结合 (NHEJ) 系统.
- 这种细菌NHEJ系统拥有DNA双链断裂 (DSB) 识别,处理和结合所需的所有活动.
- 这些发现揭示了DSB修复在不同生活领域的保存机制.
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