Kimura S, Ishibashi T, Hatanaka M, Sakakibara Y, Hashimoto J, Sakaguchi K
Department of Applied Biological Science, Faculty of Science and Technology, Science University of Tokyo, 2641 Yamazaki, Noda-shi, 278-8510, Chiba-ken, Japan
Plant Sci. 2000 Sep 8;158(1-2):33-39. doi: 10.1016/s0168-9452(00)00297-1.
By using the rice EST database, we have isolated a 2.8 kb cDNA, termed Oryza sativa ORC1 (OsORC1), from rice (O. sativa) encoding a protein that shows homology with the eukaryotic ORC1 proteins. Alignment of the OsORC1 protein sequence with the sequence of ORC1 from human and yeasts S. cerevisiae and S. pombe showed a high degree of sequence homology (38.7, 32.9 and 35.0% identity, respectively), particularly around the C-terminal region containing the CDC-NTP domain. Interestingly, the OsORC1 protein had an A+T hook-like motif, which was not present in the human or yeast genes. Genomic analysis indicated that OsORC1 existed as a single copy per genome. OsORC1 transcripts were expressed strongly in root tips and weakly in young leaves containing root apical meristem and marginal meristem, respectively. No expression was detected in the mature leaves. The level of OsORC1 expression was significantly reduced when cell proliferation was temporarily halted by the removal of sucrose from the growth medium. When the growth-halted cells began to re-grow following addition of sucrose to the medium, OsORC1 was again expressed at high levels. These results suggested that OsORC1 is required for cell proliferation. The role of OsORC1 in plant DNA replication will be discussed.
通过利用水稻EST数据库,我们从水稻(O. sativa)中分离出了一个2.8 kb的cDNA,命名为水稻ORC1(OsORC1),它编码一种与真核生物ORC1蛋白具有同源性的蛋白质。将OsORC1蛋白序列与人、酿酒酵母和粟酒裂殖酵母的ORC1序列进行比对,结果显示出高度的序列同源性(分别为38.7%、32.9%和35.0%的一致性),特别是在包含CDC-NTP结构域的C末端区域附近。有趣的是,OsORC1蛋白具有一个A+T钩样基序,而在人类或酵母基因中不存在。基因组分析表明,OsORC1在每个基因组中以单拷贝形式存在。OsORC1转录本在根尖中强烈表达,而在分别含有根尖分生组织和边缘分生组织的幼叶中表达较弱。在成熟叶中未检测到表达。当通过从生长培养基中去除蔗糖使细胞增殖暂时停止时,OsORC1的表达水平显著降低。当向培养基中添加蔗糖后生长停止的细胞开始重新生长时,OsORC1再次高水平表达。这些结果表明,OsORC1是细胞增殖所必需的。将讨论OsORC1在植物DNA复制中的作用。