Carty M P, Zernik-Kobak M, McGrath S, Dixon K
Department of Environmental Health, College of Medicine, University of Cincinnati, OH 45267-0056.
EMBO J. 1994 May 1;13(9):2114-23. doi: 10.1002/j.1460-2075.1994.tb06487.x.
We show that DNA replication activity in extracts of human HeLa cells decreases following UV irradiation. Alterations in replication activity in vitro parallel the UV-induced block in cell cycle progression of these cells in culture. UV irradiation also induces specific changes in the pattern of phosphorylation of the 34 kDa subunit of a DNA replication protein, human single-stranded DNA-binding protein (hSSB). The appearance of a hyperphosphorylated form of hSSB correlates with reduced in vitro DNA replication activity in extracts of UV-irradiated cells. Replication activity can be restored to these extracts in vitro by addition of purified hSSB. These results suggest that UV-induced DNA synthesis arrest may be mediated in part through phosphorylation-related alterations in the activity of hSSB, an essential component of the DNA replication apparatus.
我们发现,人HeLa细胞提取物中的DNA复制活性在紫外线照射后会降低。体外复制活性的改变与这些培养细胞中紫外线诱导的细胞周期进程阻滞情况相似。紫外线照射还会诱导DNA复制蛋白人单链DNA结合蛋白(hSSB)的34 kDa亚基磷酸化模式发生特定变化。hSSB超磷酸化形式的出现与紫外线照射细胞提取物中体外DNA复制活性降低相关。通过添加纯化的hSSB可在体外将这些提取物的复制活性恢复。这些结果表明,紫外线诱导的DNA合成停滞可能部分是通过DNA复制装置的重要组成部分hSSB活性的磷酸化相关改变介导的。