A Holliday junction resolvase from Pyrococcus furiosus:: Functional similarity to Escherichia coli RuvC provides evidence for conserved mechanism of homologous recombination in Bacteria, Eukarya, and Archaea
A Holliday junction resolvase from Pyrococcus furiosus:: Functional similarity to Escherichia coli RuvC provides evidence for conserved mechanism of homologous recombination in Bacteria, Eukarya, and Archaea
复制标题
DOI：
10.1073/pnas.96.16.8873
复制
发表时间：
1999-08-03
影响因子：
11.1
通讯作者：
Ishino, Y
中科院分区：
文献类型：
--
作者：
Komori, K;Sakae, S;Ishino, Y
Holliday连接蛋白是同源重组的重要中间体，细菌的RecA、真核生物的Rad51和酵母的RadA是结构和功能上的同源蛋白，它们在两个同源DNA双链体形成Holliday连接蛋白中起着关键作用。RuvC是一种特异性核酸内切酶，可分解细菌中的霍利迪连接。已经在酵母和哺乳动物细胞中发现了霍利迪连接解析活性。为了检查同源重组的范例是否适用于酵母，我们测定并发现了在激烈火球菌细胞的粗提取物中解析合成霍利迪连接的活性。该基因hjc（Holliday junction cleavage）编码由123个氨基酸组成的蛋白质，其序列与任何已知功能的蛋白质的序列都不相似。然而，所有四个古生菌，其全基因组序列已公布，具有同源基因。纯化的Njc蛋白在体外切割RecA形成的重组中间体。这些结果支持了Holliday连接的形成和分解是三个生命领域同源重组的共同机制的观点。
The Holliday junction is an essential intermediate of homologous recombination, RecA of Bacteria, Rad51 of Eukarya, and RadA of Archaea are structural and functional homologs, These proteins play a pivotal role in the formation of Holliday junctions from two homologous DNA duplexes. RuvC is a specific endonuclease that resolves Holliday junctions in Bacteria. A Holliday junction-resolving activity has been found in both yeast and mammalian cells, To examine whether the paradigm of homologous recombination apply to Archaea, we assayed and found the activity to resolve a synthetic Holliday junction in crude extract of Pyrococcus furiosus cells. The gene, hjc (Holliday junction cleavage), encodes a protein composed of 123 amino acids, whose sequence is not similar to that of any proteins with known function. However, all four archaea, whose total genome sequences have been published, have the homologous genes. The purified Njc protein cleaved the recombination intermediates formed by RecA in vitro. These results support the notion that the formation and resolution of Holliday junction is the common mechanism of homologous recombination in the three domains of life.