Lehner C F, O'Farrell P H
Department of Biochemistry and Biophysics, University of California, San Francisco 94143.
EMBO J. 1990 Nov;9(11):3573-81. doi: 10.1002/j.1460-2075.1990.tb07568.x.
Using probes obtained by PCR amplification, we have cloned Drosophila cDNAs encoding structural homologs of the p34cdc2 cell cycle kinase. Southern blot experiments and in situ hybridization to polytene chromosomes demonstrated that the isolated cDNAs, were derived from two distinct genes, Dm cdc2 (31E) and Dm cdc2c (92F). Northern blot and in situ hybridization experiments revealed that these two genes are coexpressed during embryogenesis and that expression is correlated with cell proliferation. However, despite the similarity in structure and expression, the two gene products differed in functional assays in yeasts. Expression of Dm cdc2 in Schizosaccharomyces pombe and Saccharomyces cerevisiae rescued cell cycle arrest caused by mutations in cdc2+ and CDC28, the genes encoding the p34cdc2 kinase homologs of these yeasts. In contrast, the Dm cdc2c gene product did not restore cell cycle progression. Thus, in addition to the identification of a functional homolog in Drosophila, our results indicate the presence of a closely related cognate of the p34cdc2 cell cycle kinase.
利用通过聚合酶链反应(PCR)扩增获得的探针,我们克隆了果蝇中编码p34cdc2细胞周期激酶结构同源物的cDNA。Southern印迹实验以及对多线染色体的原位杂交表明,分离得到的cDNA来自两个不同的基因,即Dm cdc2(31E)和Dm cdc2c(92F)。Northern印迹和原位杂交实验显示,这两个基因在胚胎发育过程中共同表达,且表达与细胞增殖相关。然而,尽管结构和表达相似,但这两种基因产物在酵母功能测定中存在差异。Dm cdc2在粟酒裂殖酵母和酿酒酵母中的表达挽救了由cdc2 +和CDC28突变导致的细胞周期停滞,这两个基因分别编码这些酵母的p34cdc2激酶同源物。相反,Dm cdc2c基因产物未能恢复细胞周期进程。因此,除了在果蝇中鉴定出一种功能同源物外,我们的结果还表明存在一种与p34cdc2细胞周期激酶密切相关的同源物。