Hashimoto J, Hirabayashi T, Hayano Y, Hata S, Ohashi Y, Suzuka I, Utsugi T, Toh-e A, Kikuchi Y
National Institute of Agrobiological Resources, Ibaraki, Japan.
Mol Gen Genet. 1992 May;233(1-2):10-6. doi: 10.1007/BF00587555.
Using probes obtained by PCR amplification, we have isolated two cognate rice cDNAs (cdc2Os-1 and cdc2Os-2) encoding structural homologues of the cdc2+/CDC28 (cdc2) protein kinase from a cDNA library prepared from cultured rice cells. Comparison of the deduced amino acid sequences of cdc2Os-1 and cdc2Os-2 showed that they are 83% identical. They are 62% identical to CDC28 of Saccharomyces cerevisiae and much more similar to the yeast and mammalian p34cdc2 kinases than to rice R2, a cdc2-related kinase isolated previously by screening the same rice cDNA library with a different oligonucleotide probe. Southern blot analysis indicated that the three rice clones (cdc2Os-1, cdc2Os-2 and R2) are derived from distinct genes and are each found in a single copy per rice haploid genome. RNA blot analysis revealed that these genes are expressed in proliferating rice cells and in young rice seedlings. cdc2Os-1 could complement a temperature-sensitive yeast mutant of cdc28. However, despite the similarity in structure, both cdc2Os-2 and R2 were unable to complement the same mutant. Thus, the present results demonstrate the presence of structurally related, but functionally distinct cognates of the cdc2 cell cycle kinase in rice.
利用通过聚合酶链式反应(PCR)扩增获得的探针,我们从培养的水稻细胞制备的cDNA文库中分离出了两个同源的水稻cDNA(cdc2Os - 1和cdc2Os - 2),它们编码cdc2⁺/CDC28(cdc2)蛋白激酶的结构同源物。对cdc2Os - 1和cdc2Os - 2推导的氨基酸序列进行比较,结果表明它们的同源性为83%。它们与酿酒酵母的CDC28的同源性为62%,并且与酵母和哺乳动物的p34cdc2激酶相比,与水稻R2(先前用不同的寡核苷酸探针筛选同一水稻cDNA文库分离得到的一种与cdc2相关的激酶)更为相似。Southern印迹分析表明,这三个水稻克隆(cdc2Os - 1、cdc2Os - 2和R2)源自不同的基因,并且在每个水稻单倍体基因组中均以单拷贝形式存在。RNA印迹分析显示,这些基因在增殖的水稻细胞和水稻幼苗中表达。cdc2Os - 1能够互补cdc28的温度敏感型酵母突变体。然而,尽管在结构上相似,但cdc2Os - 2和R2均无法互补同一突变体。因此,目前的结果表明水稻中存在结构相关但功能不同的cdc2细胞周期激酶同源物。