Atomic Force Microscopic Study of Low Temperature Induced Disassembly of RecA-dsDNA Filaments
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- 作者：Cunlan Guo ; Yonghai Song ; Li Wang ; Lanlan Sun ; Yujing Sun ; Chongyang Peng ; Zhelin Liu ; Tao Yang ; Zhuang Li
- 刊名：Journal of Physical Chemistry B
- 出版年：2008
- 出版时间：January 24, 2008
- 年：2008
- 卷：112
- 期：3
- 页码：1022 - 1027
- 全文大小：328K
- 年卷期：v.112,no.3(January 24, 2008)
- ISSN：1520-5207
文摘
The assembly and disassembly of RecA-DNA nucleoprotein filaments on double-stranded DNA (dsDNA)or single-stranded DNA (ssDNA) are important steps for homologous recombination and DNA repair. Theassembly and disassembly of the nucleoprotein filaments are sensitive to the reaction conditions. In this work,we investigated different morphologies of the formed nucleoprotein filaments at low temperature under differentsolution conditions by atomic force microscopy (AFM). We found that low temperature and long keepingtime could induce the incomplete disassembly of the formed nucleoprotein filaments. In addition, when theformed filaments were kept at -20 
C for 20 h with 1,4-dithiothreitol (DTT), the integrated filamentsdisassembled. It was similar to the case under the same condition without anything added. However, whenglycerol was used as a substitute for DTT, there was no obvious disassembly at the same condition. Oppositely,when the formed filaments were kept at 4 C for 20 h, the disassembly with additional DTT was not asobvious as the case at -20 C for 20 h, whereas the case with additional glycerol disassembled. The experimentsindicated the effect of cold denaturation on the interaction of DNA and RecA. Meanwhile, the study of thesephenomena can supply guidelines for the property and stability of RecA as well as the relevant roles ofinfluencing factors to RecA and DNA in further theoretical studies.
C for 20 h with 1,4-dithiothreitol (DTT), the integrated filamentsdisassembled. It was similar to the case under the same condition without anything added. However, whenglycerol was used as a substitute for DTT, there was no obvious disassembly at the same condition. Oppositely,when the formed filaments were kept at 4 C for 20 h, the disassembly with additional DTT was not asobvious as the case at -20 C for 20 h, whereas the case with additional glycerol disassembled. The experimentsindicated the effect of cold denaturation on the interaction of DNA and RecA. Meanwhile, the study of thesephenomena can supply guidelines for the property and stability of RecA as well as the relevant roles ofinfluencing factors to RecA and DNA in further theoretical studies.