The nuclease domain of adeno-associated virus rep coordinates replication initiation using two distinct DNA recognition interfaces
The nuclease domain of adeno-associated virus rep coordinates replication initiation using two distinct DNA recognition interfaces
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DOI：
10.1016/s1097-2765(04)00023-1
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发表时间：
2004-02-13
期刊：
影响因子：
16
通讯作者：
Dyda, F
中科院分区：
文献类型：
--
作者：
Hickman, AB;Ronning, DR;Dyda, F
整合到人类染色体的特定位置是腺相关病毒（AAV）的独特性质。该反应需要病毒Rep蛋白和AAV起始序列。为了了解Rep如何识别DNA，我们已经确定了Rep核酸内切酶结构域分别与两种DNA底物复合的结构：病毒反向末端重复序列内的Rep结合位点和末端发夹臂之一。在Rep结合位点，五个Rep单体结合五个四核苷酸直接重复-每个重复被两个Rep单体从DNA的相对面识别。茎-环结合涉及在分子的与ssDNA被切割的活性位点相对的一侧上的蛋白质界面。因此，Rep在其核酸内切酶结构域内具有三个不同的结合位点，用于其不同的DNA底物。使用这些不同的界面产生了调节病毒复制和整合后期事件所必需的结构不对称性。
Integration into a particular location in human chromosomes is a unique property of the adeno-associated virus (AAV). This reaction requires the viral Rep protein and AAV origin sequences. To understand how Rep recognizes DNA, we have determined the structures of the Rep endonuclease domain separately complexed with two DNA substrates: the Rep binding site within the viral inverted terminal repeat and one of the terminal hairpin arms. At the Rep binding site, five Rep monomers bind five tetranucleotide direct repeats-each repeat is recognized by two Rep monomers frog opposing faces of the DNA. Stem-loop binding involves a protein interface on the opposite side of the molecule from the active site where ssDNA is cleaved. Rep therefore has three distinct binding sites within its endonuclease domain for its different DNA substrates. Use of these different interfaces generates the structural asymmetry necessary to regulate later events in viral replication and integration.