Homologous pairing activities of two rice RAD51 proteins, RAD51A1 and RAD51A2.
Homologous pairing activities of two rice RAD51 proteins, RAD51A1 and RAD51A2.
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DOI：
10.1371/journal.pone.0075451
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Kurumizaka H
中科院分区：
文献类型：
--
作者：
Morozumi Y;Ino R;Ikawa S;Mimida N;Shimizu T;Toki S;Ichikawa H;Shibata T;Kurumizaka H
在高等真核生物中，RAD 51作为同源重组和DNA双链断裂的重组修复中的必需蛋白发挥功能。在这些过程中，RAD51催化单链DNA和双链DNA之间的同源配对。水稻（Oryza sativa）的粳稻栽培品种编码两个RAD 51蛋白，RAD 51A1和RAD 51A2，而酵母和哺乳动物中仅存在一个RAD 51。然而，RAD51A1和RAD51A2之间的功能差异尚未阐明，因为它们的生化特性尚未被表征。在本研究中，我们纯化了RAD51A1和RAD51A2，并发现RAD51A2在体外强烈促进同源配对。RAD51A1也具有同源配对活性，但其活性仅为RAD51A2活性的约10%。RAD51A1和RAD51A2都与ssDNA和dsDNA结合，它们的DNA结合严格需要ATP，ATP调节RAD51A1和RAD51A2的聚合物形成活性。这些结果表明，尽管RAD51A1和RAD51A2都具有催化同源配对的潜力，但RAD51A2可能是水稻中的主要重组酶。
In higher eukaryotes, RAD51 functions as an essential protein in homologous recombination and recombinational repair of DNA double strand breaks. During these processes, RAD51 catalyzes homologous pairing between single-stranded DNA and double-stranded DNA. Japonica cultivars of rice (Oryza sativa) encode two RAD51 proteins, RAD51A1 and RAD51A2, whereas only one RAD51 exists in yeast and mammals. However, the functional differences between RAD51A1 and RAD51A2 have not been elucidated, because their biochemical properties have not been characterized. In the present study, we purified RAD51A1 and RAD51A2, and found that RAD51A2 robustly promotes homologous pairing in vitro. RAD51A1 also possesses homologous-pairing activity, but it is only about 10% of the RAD51A2 activity. Both RAD51A1 and RAD51A2 bind to ssDNA and dsDNA, and their DNA binding strictly requires ATP, which modulates the polymer formation activities of RAD51A1 and RAD51A2. These findings suggest that although both RAD51A1 and RAD51A2 have the potential to catalyze homologous pairing, RAD51A2 may be the major recombinase in rice.
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