Interactions between UvrA and UvrB: the role of UvrB's domain 2 in nucleotide excision repair
Interactions between UvrA and UvrB: the role of UvrB's domain 2 in nucleotide excision repair
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DOI：
10.1038/sj.emboj.7600263
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发表时间：
2004-07-07
期刊：
影响因子：
11.4
通讯作者：
Kisker, C
中科院分区：
文献类型：
--
作者：
Truglio, JJ;Croteau, DL;Kisker, C
核苷酸切除修复（NER）是一种高度保守的DNA修复机制，存在于所有生物王国中。UvrB是细菌NER系统的核心组成部分，参与损伤识别、链切除和修复合成。目前已知的三种UvrB晶体结构都没有定义域2的结构，而域2是与UvrB相互作用的关键。我们已经解决了UvrB Y96A变体的晶体结构，揭示了结构域2的新折叠，并确定了位于其表面的高度保守残基。这些残基局限于对DNA结合重要的UvrB表面，可能对UvrB与UvrA的相互作用至关重要。我们对这些残基进行了突变，以研究它们在切割反应、切割前复合物的形成、DNA短双工区域的不稳定、与UvrA的结合和ATP水解中的作用。基于结构和生化数据，我们得出结论，结构域2是有效的UvrA-UvrB相互作用所必需的，这是核苷酸切除修复所有后续步骤的先决条件。
Nucleotide excision repair (NER) is a highly conserved DNA repair mechanism present in all kingdoms of life. UvrB is a central component of the bacterial NER system, participating in damage recognition, strand excision and repair synthesis. None of the three presently available crystal structures of UvrB has defined the structure of domain 2, which is critical for the interaction with UvrA. We have solved the crystal structure of the UvrB Y96A variant, which reveals a new fold for domain 2 and identifies highly conserved residues located on its surface. These residues are restricted to the face of UvrB important for DNA binding and may be critical for the interaction of UvrB with UvrA. We have mutated these residues to study their role in the incision reaction, formation of the pre-incision complex, destabilization of short duplex regions in DNA, binding to UvrA and ATP hydrolysis. Based on the structural and biochemical data, we conclude that domain 2 is required for a productive UvrA-UvrB interaction, which is a pre-requisite for all subsequent steps in nucleotide excision repair.