SWI5-SFR1與RAD51AP1對RAD51重組酵素的作用機轉

廖嘉妤

doi:10.6342/NTU.2013.01899

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SWI5-SFR1与RAD51AP1对RAD51重组酵素的作用机转

The mechanism of SWI5-SFR1 and RAD51AP1 stimulating RAD51-mediated recombination

廖嘉妤 , 硕士　　导师：冀宏源　　

英文 DOI： 10.6342/NTU.2013.01899

同源重组；相乘作用； Rad51重组酵素； Swi5-Sfr1 ； Rad51AP1 ； Homologous recombination ； Synergistic effect ； Rad51 recombinase ； Rad51AP1 ； Swi5-Sfr1

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摘要 │ 参考文献 (47) │ 国际计量

摘要〈TOP〉

并列摘要〈TOP〉

Homologous recombination (HR) is the major pathway to repair DNA double-strand breaks (DSBs). Dysfunction of HR leads to genome instability and even cancer formation. When a double-stranded DNA (dsDNA) breaks, the break ends undergo a resection process to generate a single-stranded DNA (ssDNA), and then RAD51 recombinase assembles on ssDNA to form a nucleoprotein filament, which is called presynaptic filament. The presynaptic filament captures and invades in dsDNA to search the homologous DNA. Finally, the ssDNA exchanges with their homologous DNA within dsDNA. During this HR reaction, the activity of RAD51 is regulated by accessory factors, including SWI5-SFR1 and RAD51AP1. Previous cell-based studies showed that knockdown SWI5-SFR1 or RAD51AP1 in mammalian cells results in increased sensitivity to DNA damaging agents. It raises an intriguing question how SWI5-SFR1 and RAD51AP1 mechanistically co-regulate Rad51 recombinase activity during HR. To address this question, we use purified proteins and in vitro reconstitution system to study their mechanistic actions in recombination reaction. Our results indicate that mouse SWI5-SFR1 and RAD51AP1 exhibit a synergistic effect on RAD51-mediated strand exchange. Moreover, the synergistic effect not only requires SWI5-SFR1 forming complex interacting with RAD51, but also requires RAD51AP1 binding DNA and RAD51. Furthermore, we found that SWI5-SFR1 can’t associate with RAD51AP1 directly, but SWI5-SFR1, RAD51AP1 and RAD51 form a ternary complex. Taken together, our findings suggest that the synergistic effect stems from the stimulation of SWI5-SFR1 and RAD51AP1 individually, and provide a mechanical insight of accessory factors in homologous recombination.

参考文献 ( 47 ) 〈TOP〉

1. Akamatsu, Y., Dziadkowiec, D., Ikeguchi, M., Shinagawa, H., and Iwasaki, H. (2003). Two different Swi5-containing protein complexes are involved in mating-type switching and recombination repair in fission yeast. Proceedings of the National Academy of Sciences of the United States of America 100, 15770-15775.
连结：
2. Akamatsu, Y., and Jasin, M. (2010). Role for the mammalian Swi5-Sfr1 complex in DNA strand break repair through homologous recombination. PLoS genetics 6, e1001160.
连结：
3. Akamatsu, Y., Tsutsui, Y., Morishita, T., Siddique, M.S., Kurokawa, Y., Ikeguchi, M., Yamao, F., Arcangioli, B., and Iwasaki, H. (2007). Fission yeast Swi5/Sfr1 and Rhp55/Rhp57 differentially regulate Rhp51-dependent recombination outcomes. The EMBO journal 26, 1352-1362.
连结：
4. Bishop, D.K., Park, D., Xu, L., and Kleckner, N. (1992). DMC1: a meiosis-specific yeast homolog of E. coli recA required for recombination, synaptonemal complex formation, and cell cycle progression. Cell 69, 439-456.
连结：
6. Dunlop, M.H., Dray, E., Zhao, W., San Filippo, J., Tsai, M.S., Leung, S.G., Schild, D., Wiese, C., and Sung, P. (2012). Mechanistic insights into RAD51-associated protein 1 (RAD51AP1) action in homologous DNA repair. The Journal of biological chemistry 287, 12343-12347.
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引用含有DOI的文献

DOI可强化引用精确性、增强学术圈连结，并给予使用者跨平台的良好使用经验，目前在全世界已有超过五千万个对象申请DOI。 如想对DOI的使用与概念有进一步了解，请参考 （ ） 。

DOI可强化引用精确性、增强学术圈连结，并给予使用者跨平台的良好使用经验，目前在全世界已有超过五千万个对象申请DOI。如想对DOI的使用与概念有进一步了解，请参考（）。