Investigate the Biochemical Characteristic of BCDX2 Complex and Its Functional Role with RAD51
| Autor: | Yi-Zhen Jiang, 江宜蓁 |
|---|---|
| Rok vydání: | 2017 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 106 Homologous recombination (HR) plays a central role in maintaining genomic integrity by repairing DNA double-strand breaks (DSBs). The evolutionarily conserved RAD51 recombinase is the key enzyme to execute the recombination-mediated DSB repair. Moreover, cell-based studies have well documented that five RAD51 paralogs forming two distinct complexes BCDX2 and CX3 are required for RAD51 activity. These RAD51 paralogs share about 20-30% sequence identity at amino acid level with RAD51 and possess the ability of ATP binding and hydrolysis. However, it remains largely unknown about the mechanistic perspective of the BCDX2 complex in regulating RAD51 activity due to the difficulty of obtaining BCDX2 protein complex. Here we successfully established expression and purification procedures of the BCDX2 complex in the human expression system. The purified BCDX2 complex exhibits the single-strand DNA binding activity and physically interacts with RAD51. To our surprise, we found that RAD51B could be phosphorylated in vitro. Our preliminary data indicates that the phosphorylation of RAD51B affects the protein-protein interaction between BCDX2 complex and RAD51 recombinase. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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