Hsieh Hui-Chuan, Hsieh Yi-Hsuan, Huang Yu-Hsin, Shen Fan-Ching, Tsai Han-Ni, Tsai Jui-He, Lai Yu-Ting, Wang Yu-Ting, Chuang Woei-Jer, Huang Wenya
Medical Laboratory Science and Biotechnology, College of Medicine, National Cheng Kung University, Tainan 70101, Taiwan.
Biochem Biophys Res Commun. 2005 Sep 16;335(1):181-7. doi: 10.1016/j.bbrc.2005.07.067.
HHR23A and hHR23B are the human homologs of Saccharomyces cerevisiae Rad23. hHR23B is associated with the nucleotide excision repair (NER) factor xeroderma pigmentosum C (XPC) protein and is required for global genome repair. The function of hHR23A is not yet clear. In this study, the potential function of the hHR23A protein was investigated using RNA interference techniques. The hHR23A knock-down (KD) construct diminished the RNA level of hHR23A protein by approximately 60%, and it did not interfere with expression of the hHR23B gene. Based on Southwestern immunoblot and host-cell reactivation assays, hHR23A(KD) cells were found to be deficient in DNA repair activity against the DNA damage caused by UVC irradiation. In these hHR23A(KD) cells, the XPC gene was not normally induced by UVC irradiation, indicating that the hHR23A protein is involved in NER through regulation of the DNA damage recognition protein XPC. Co-immunoprecipitation experiments revealed that hHR23A was associated with a small portion of hHR23B and the majority of p53 protein, indicating that hHR23A regulates the function of XPC by its association with the NER activator p53.
HHR23A和hHR23B是酿酒酵母Rad23的人类同源物。hHR23B与核苷酸切除修复(NER)因子着色性干皮病C(XPC)蛋白相关,是全基因组修复所必需的。hHR23A的功能尚不清楚。在本研究中,使用RNA干扰技术研究了hHR23A蛋白的潜在功能。hHR23A敲低(KD)构建体使hHR23A蛋白的RNA水平降低了约60%,且不干扰hHR23B基因的表达。基于蛋白质免疫印迹和宿主细胞再激活试验,发现hHR23A(KD)细胞对紫外线(UVC)照射引起的DNA损伤缺乏DNA修复活性。在这些hHR23A(KD)细胞中,UVC照射不能正常诱导XPC基因,这表明hHR23A蛋白通过调节DNA损伤识别蛋白XPC参与核苷酸切除修复。免疫共沉淀实验表明,hHR23A与一小部分hHR23B和大部分p53蛋白相关,这表明hHR23A通过与核苷酸切除修复激活剂p53结合来调节XPC的功能。