The role of KIAA0101 in cell cycle progression and DNA repair.
| Autor: | Mow-Jung Feng, 酆茂蓉 |
|---|---|
| Rok vydání: | 2010 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 98 By microarray analysis, we identified KIAA0101 (p15PAF) as a gene overexpressed in hepatocellular carcinoma. KIAA0101 is a proliferating cell nuclear antigen (PCNA)-binding protein of unknown function. In this study, we found that KIAA0101 was expressed only in S phase of the cell cycle. Knockdown of KIAA0101 inhibited cell cycle progression from G1 to S phase and suppressed BrdU incorporation. Knockdown of KIAA0101 induced p21 overexpression. To further delineate the molecular mechanism for the function of KIAA0101 protein, the yeast two hybrid assay was used to identify the interacting proteins. Rad23A was identified as a putative KIAA0101-binding protein. Using the immunoprecipitation assay, we demonstrated that EGFP-tagged KIAA0101 interacted with Rad23A, confirming the interaction of KIAA0101 and Rad23A in vivo. Rad23A is a major player in nucleotide excision repair. This result suggests that KIAA0101 play a role in DNA damage repair. Consistent with this suggestion, colony formation assay showed that KIAA0101 depletion sensitized cells to UV. Moreover, DNA synthesis was decreased in cells with KIAA0101 knockdown under UV irradiation. These results suggest that KIAA0101 is a multifunctional PCNA-interacting protein, and play important roles in regulating DNA synthesis and repair. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
| Externí odkaz: |
|