Hirayama T, Shinozaki K
Laboratory of Plant Molecular Biology, Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), Ibaraki, Japan.
Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):13371-6. doi: 10.1073/pnas.93.23.13371.
We cloned and characterized a cDNA corresponding to a cdc5+ homolog of the higher plant, Arabidopsis thaliana. The cDNA, named AtCDC5 cDNA, encodes a polypeptide of 844 amino acid residues. The amino acid sequence of N-terminal one-fourth region of the predicted protein bears significant similarity to that of Schizosaccharomyces pombe Cdc5 and Myb-related proteins. Overexpression of the AtCDC5 cDNA in S. pombe cells is able to complement the growth defective phenotype of a cdc5 temperature-sensitive mutant. These results indicate that the AtCDC5 gene is a plant counterpart of S. pombe cdc5+. This is the first report of a cdc5(+)-like gene in a multicellular organism. We also demonstrated that a recombinant AtCDC5 protein possesses a sequence specific DNA binding activity (CTCAGCG) and the AtCDC5 gene is expressed extensively in shoot and root meristems. In addition, we cloned a PCR fragment corresponding to the DNA binding domain of human Cdc5-like protein. These results strongly suggest that Cdc5-like protein exists in all eukaryotes and may function in cell cycle regulation.
我们克隆并鉴定了与高等植物拟南芥的cdc5 +同源物对应的cDNA。该cDNA名为AtCDC5 cDNA,编码一个由844个氨基酸残基组成的多肽。预测蛋白N端四分之一区域的氨基酸序列与粟酒裂殖酵母Cdc5和Myb相关蛋白的氨基酸序列具有显著相似性。AtCDC5 cDNA在粟酒裂殖酵母细胞中的过表达能够弥补cdc5温度敏感突变体的生长缺陷表型。这些结果表明AtCDC5基因是粟酒裂殖酵母cdc5 +的植物对应物。这是多细胞生物中cdc5(+)样基因的首次报道。我们还证明了重组AtCDC5蛋白具有序列特异性DNA结合活性(CTCAGCG),并且AtCDC5基因在茎尖和根尖分生组织中广泛表达。此外,我们克隆了与人类Cdc5样蛋白的DNA结合结构域对应的PCR片段。这些结果强烈表明Cdc5样蛋白存在于所有真核生物中,并且可能在细胞周期调控中发挥作用。