Boxem M, Srinivasan D G, van den Heuvel S
Massachusetts General Hospital Cancer Center, Building 149, Charlestown, MA 02129, USA.
Development. 1999 May;126(10):2227-39. doi: 10.1242/dev.126.10.2227.
We have identified six protein kinases that belong to the family of cdc2-related kinases in Caenorhabditis elegans. Results from RNA interference experiments indicate that at least one of these kinases is required for cell-cycle progression during meiosis and mitosis. This kinase, encoded by the ncc-1 gene, is closely related to human Cdk1/Cdc2, Cdk2 and Cdk3 and yeast CDC28/cdc2(+). We addressed whether ncc-1 acts to promote passage through a single transition or multiple transitions in the cell cycle, analogous to Cdks in vertebrates or yeasts, respectively. We isolated five recessive ncc-1 mutations in a genetic screen for mutants that resemble larval arrested ncc-1(RNAi) animals. Our results indicate that maternal ncc-1 product is sufficient for embryogenesis, and that zygotic expression is required for cell divisions during larval development. Cells that form the postembryonic lineages in wild-type animals do not enter mitosis in ncc-1 mutants, as indicated by lack of chromosome condensation and nuclear envelope breakdown. However, progression through G1 and S phase appears unaffected, as revealed by expression of ribonucleotide reductase, incorporation of BrdU and DNA quantitation. Our results indicate that C. elegans uses multiple Cdks to regulate cell-cycle transitions and that ncc-1 is the C. elegans ortholog of Cdk1/Cdc2 in other metazoans, required for M phase in meiotic as well as mitotic cell cycles.
我们已经在秀丽隐杆线虫中鉴定出六种属于细胞周期蛋白依赖性激酶2(cdc2)相关激酶家族的蛋白激酶。RNA干扰实验结果表明,这些激酶中至少有一种在减数分裂和有丝分裂期间的细胞周期进程中是必需的。这种由ncc-1基因编码的激酶与人类细胞周期蛋白依赖性激酶1(Cdk1)/细胞周期蛋白依赖性激酶2(Cdc2)、细胞周期蛋白依赖性激酶2(Cdk2)和细胞周期蛋白依赖性激酶3(Cdk3)以及酵母CDCCDC1酵母细胞周期蛋白依赖性激酶28(CDC28)/细胞周期蛋白依赖性激酶2(cdc2(+))密切相关。我们探讨了ncc-1是像脊椎动物中的细胞周期蛋白依赖性激酶那样促进细胞周期单次转变,还是像酵母中的细胞周期蛋白依赖性激酶那样促进多次转变。我们在一项遗传筛选中分离出五个隐性ncc-1突变体,这些突变体类似于幼虫期停滞的ncc-1(RNA干扰)动物。我们的结果表明,母源ncc-1产物足以支持胚胎发育,而合子表达对于幼虫发育期间的细胞分裂是必需的。如缺乏染色体凝聚和核膜破裂所示,野生型动物中形成胚后谱系的细胞在ncc-1突变体中不会进入有丝分裂。然而,如核糖核苷酸还原酶的表达、5-溴脱氧尿嘧啶核苷(BrdU)的掺入和DNA定量分析所示,细胞通过G1期和S期的进程似乎未受影响。我们的结果表明,秀丽隐杆线虫利用多种细胞周期蛋白依赖性激酶来调节细胞周期转变,并且ncc-1是其他后生动物中细胞周期蛋白依赖性激酶1(Cdk1)/细胞周期蛋白依赖性激酶2(Cdc2)在秀丽隐杆线虫中的直系同源物,在减数分裂以及有丝分裂细胞周期的M期是必需的。