Hofmann J F, Beach D
Howard Hughes Medical Institute, Cold Spring Harbor Laboratory, NY 11724.
EMBO J. 1994 Jan 15;13(2):425-34. doi: 10.1002/j.1460-2075.1994.tb06277.x.
We have used an immunoprecipitation-PCR cycle to isolate physically genomic DNA sequences that are bound by the fission yeast cdc10 gene product in an attempt to identify novel target genes. An essential gene, cdt1, has been isolated whose expression is cell cycle regulated in a cdc10 dependent manner. The cdt1 promoter contains a recognition site for a sequence specific DNA binding factor. The cdc10 gene product is a component of this factor. Ectopic expression of cdt1 can complement a temperature sensitive mutation of cdc10 at semipermissive temperature. Cells carrying a null allele of cdt1 are defective in DNA replication but initiate mitotic events, suggesting that cdt1 is essential for the normal dependency relationship of S-phase and mitosis.
我们利用免疫沉淀 - PCR 循环从物理上分离出与裂殖酵母 cdc10 基因产物结合的基因组 DNA 序列,试图鉴定新的靶基因。一个必需基因 cdt1 已被分离出来,其表达以 cdc10 依赖的方式受到细胞周期调控。cdt1 启动子包含一个序列特异性 DNA 结合因子的识别位点。cdc10 基因产物是该因子的一个组成部分。cdt1 的异位表达可以在半允许温度下弥补 cdc10 的温度敏感突变。携带 cdt1 无效等位基因的细胞在 DNA 复制方面存在缺陷,但能启动有丝分裂事件,这表明 cdt1 对于 S 期和有丝分裂的正常依赖关系至关重要。