Narasimhachar Yadushyla, Coué Martine
Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center, Lubbock, Texas 79430, USA.
J Biol Chem. 2009 Oct 2;284(40):27235-42. doi: 10.1074/jbc.M109.008854. Epub 2009 Aug 5.
During the mitotic cell cycle, Geminin can act both as a promoter and inhibitor of initiation of DNA replication. As a promoter, Geminin stabilizes Cdt1 and facilitates its accumulation leading to the assembly of the pre-replication complex on DNA. As an inhibitor, Geminin prevents Cdt1 from loading the mini-chromosome maintenance complex onto pre-replication complexes in late S, G(2), and M phases. Here we show that during meiosis Geminin functions as a stabilizer of Cdt1 promoting its accumulation for the early division cycles of the embryo. Depletion of Geminin in Xenopus immature oocytes leads to a decrease of Cdt1 protein levels during maturation and after activation of these oocytes. Injection of exogenous recombinant Geminin into the depleted oocytes rescues Cdt1 levels demonstrating that Geminin stabilizes Cdt1 during meiosis and after fertilization. Furthermore, Geminin-depleted oocytes did not replicate their DNA after meiosis I indicating that Geminin does not act as an inhibitor of initiation of DNA replication between meiosis I and meiosis II.
在有丝分裂细胞周期中,Geminin既可以作为DNA复制起始的促进因子,也可以作为抑制因子。作为促进因子,Geminin能稳定Cdt1并促进其积累,从而导致前复制复合体在DNA上的组装。作为抑制因子,Geminin可防止Cdt1在S期晚期、G2期和M期将微小染色体维持复合体加载到前复制复合体上。在此我们表明,在减数分裂过程中,Geminin作为Cdt1的稳定剂,促进其积累以支持胚胎早期分裂周期。非洲爪蟾未成熟卵母细胞中Geminin的缺失会导致这些卵母细胞成熟过程中以及激活后Cdt1蛋白水平下降。将外源性重组Geminin注射到缺失Geminin的卵母细胞中可恢复Cdt1水平,这表明Geminin在减数分裂期间和受精后能稳定Cdt1。此外,缺失Geminin的卵母细胞在减数分裂I后不会复制其DNA,这表明Geminin在减数分裂I和减数分裂II之间并非作为DNA复制起始的抑制因子。