Chevalier S, Couturier A, Chartrain I, Le Guellec R, Beckhelling C, Le Guellec K, Philippe M, Ford C C
Département de Biologie et Génétique du développement, CNRS URA 256, Université de Rennes I, France.
J Cell Sci. 1996 Jun;109 ( Pt 6):1173-84. doi: 10.1242/jcs.109.6.1173.
The capacity to initiate DNA replication appears during oocyte maturation in Xenopus. Initiation of S phase is driven by several components which include active cyclin/cdk complexes. We have identified three Xenopus cyclin E clones showing 59% amino acid identity with human cyclin E. The recruitment of cyclin E mRNA, like cdk2 mRNA, into the polysomal fraction during oocyte maturation, results in the accumulation of the corresponding proteins in unfertilized eggs. Cyclin E mRNA remains polyadenylated during cleavage and anti-cyclin E antibodies detect Xlcyclin E in embryonic nuclei at this time. Cdk2 protein is necessary for the phosphorylation of radiolabelled cyclin E added to egg extracts. Radiolabelled Xlcyclin E enters interphase nuclei and, though stable through interphase and mitosis, is not associated with condensed mitotic chromatin. In egg extracts, endogenous Xlcyclin E rapidly associates with nuclei before S phase and remains nuclear throughout interphase, becoming nucleoplasmic in G2/prophase. Under conditions where initiation of replication is limiting in extracts, Xlcyclin E associates only with those nuclei that undergo S phase. These features are entirely consistent with the view that Xlcyclin E is required for initiation of S phase.
非洲爪蟾卵母细胞成熟过程中会出现启动DNA复制的能力。S期的启动由多种成分驱动,其中包括活性细胞周期蛋白/细胞周期蛋白依赖性激酶(cyclin/cdk)复合物。我们鉴定出了三个非洲爪蟾细胞周期蛋白E克隆,它们与人类细胞周期蛋白E的氨基酸同一性为59%。在卵母细胞成熟过程中,细胞周期蛋白E mRNA与细胞周期蛋白依赖性激酶2(cdk2)mRNA一样,被募集到多核糖体部分,导致相应蛋白质在未受精卵中积累。在卵裂过程中,细胞周期蛋白E mRNA保持多聚腺苷酸化状态,此时抗细胞周期蛋白E抗体可在胚胎细胞核中检测到非洲爪蟾细胞周期蛋白E(Xlcyclin E)。细胞周期蛋白依赖性激酶2蛋白对于添加到卵提取物中的放射性标记细胞周期蛋白E的磷酸化是必需的。放射性标记的Xlcyclin E进入间期细胞核,虽然在间期和有丝分裂过程中稳定,但不与浓缩的有丝分裂染色质结合。在卵提取物中,内源性Xlcyclin E在S期之前迅速与细胞核结合,并在整个间期保持在细胞核内,在G2期/前期变为核质。在提取物中复制起始受到限制的条件下,Xlcyclin E仅与那些进入S期的细胞核结合。这些特征完全符合Xlcyclin E是S期起始所必需的这一观点。