Fujita M, Kiyono T, Hayashi Y, Ishibashi M
Laboratory of Viral Oncology, Research Institute, Aichi Cancer Center, Chikusa-ku, Nagoya 464, Japan.
J Biol Chem. 1996 Feb 23;271(8):4349-54. doi: 10.1074/jbc.271.8.4349.
hCDC47 is a putative human homologue of yeast CDC47 and a member of the MCM protein family, which has been implicated in the regulatory machinery causing DNA to replicate only once in the S phase. In the present study, we performed an initial characterization of hCDC47. We found that hCDC47 protein was present in the nucleus of cultured human cells in two different forms; one extractable by a non-ionic detergent and the other resistant to such extraction and tightly associated with the nucleus. The levels of the nucleus-bound form gradually diminished during S phase progression, although the total amount of nuclear hCDC47 protein remained relatively constant, suggesting that the nucleus-bound form becomes dissociated from the nuclear structure during DNA replication. This behavior of hCDC47 protein is very similar to that of other mammalian MCM proteins reported recently. We also found that expression of hCDC47 mRNA was repressed in quiescent cells but was induced at the late G1 to S phase by growth factor stimulation. Together, these findings indicate that hCDC47 protein together with other MCM proteins participates in the regulation of mammalian DNA replication.
hCDC47是酵母CDC47假定的人类同源物,也是MCM蛋白家族的成员,该家族与导致DNA在S期仅复制一次的调控机制有关。在本研究中,我们对hCDC47进行了初步表征。我们发现hCDC47蛋白以两种不同形式存在于培养的人类细胞的细胞核中;一种可被非离子去污剂提取,另一种对这种提取具有抗性并与细胞核紧密结合。尽管细胞核中hCDC47蛋白的总量保持相对恒定,但在S期进程中,与细胞核结合的形式的水平逐渐降低,这表明在DNA复制过程中,与细胞核结合的形式从核结构上解离。hCDC47蛋白的这种行为与最近报道的其他哺乳动物MCM蛋白的行为非常相似。我们还发现,hCDC47 mRNA的表达在静止细胞中受到抑制,但在G1晚期至S期通过生长因子刺激而被诱导。总之,这些发现表明hCDC47蛋白与其他MCM蛋白一起参与了哺乳动物DNA复制的调控。