Kanungo J
Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta, Georgia, USA.
Folia Biol (Praha). 2009;55(6):218-23.
DNA-dependent protein kinase is a serine/ threonine kinase consisting of a catalytic subunit, p460, and a regulatory subunit called Ku (p80/p70). DNA-dependent protein kinase plays a role in transcription, non-homologous recombination, and DNA repair. Previous data have shown the presence of DNA-dependent protein kinase in Xenopus oocytes and changes in its activity during vitellogenesis. Metabolic labelling studies have shown that the increased enzyme activity in vitellogenic oocytes correlates with increased levels of Ku protein, and compared to the pre- and early-vitellogenic oocytes, vitellogenic and post-vitellogenic oocytes show an increased level of DNA-dependent protein kinase activity. Whether DNA-dependent protein kinase activity is altered during early embryogenesis in Xenopus is not known. The present study demonstrates that DNA-dependent protein kinase activity is gradually lost from the cytoplasm in the early embryonic cells of blastulae and gastrulae. The gradual loss of DNA-dependent protein kinase activity during post-fertilization early embryogenesis in Xenopus is consistent with the reports on Arbacia punctulata. Immunohistochemistry of the oocytes at various stages and early embryos (gastrulae) shows that Ku70, a regulatory subunit of DNA-dependent protein kinase, is present both in the cytoplasm and nucleus in the pre-vitellogenic oocytes, full-grown post-vitellogenic oocytes and in the gastrula cells. However, Ku70 appears to accumulate in higher concentrations in the nuclei of gastrula cells. These results suggest that gradual loss of DNA-dependent protein kinase activity from the cytoplasm of the early embryos could be the consequence of Ku translocating to the nuclei that may be necessary for post-zygotic transcription followed by cellular differentiation.
DNA依赖性蛋白激酶是一种丝氨酸/苏氨酸激酶,由催化亚基p460和称为Ku(p80/p70)的调节亚基组成。DNA依赖性蛋白激酶在转录、非同源重组和DNA修复中发挥作用。先前的数据表明非洲爪蟾卵母细胞中存在DNA依赖性蛋白激酶,并且在卵黄发生过程中其活性会发生变化。代谢标记研究表明,卵黄生成期卵母细胞中酶活性的增加与Ku蛋白水平的增加相关,与卵黄生成前期和早期卵母细胞相比,卵黄生成期和卵黄生成后期的卵母细胞显示出DNA依赖性蛋白激酶活性水平增加。非洲爪蟾早期胚胎发育过程中DNA依赖性蛋白激酶活性是否发生改变尚不清楚。本研究表明,在囊胚和原肠胚的早期胚胎细胞中,DNA依赖性蛋白激酶活性从细胞质中逐渐丧失。非洲爪蟾受精后早期胚胎发育过程中DNA依赖性蛋白激酶活性的逐渐丧失与关于斑点海胆的报道一致。对不同阶段的卵母细胞和早期胚胎(原肠胚)进行免疫组织化学分析表明,DNA依赖性蛋白激酶的调节亚基Ku70在卵黄生成前期卵母细胞、成熟的卵黄生成后期卵母细胞和原肠胚细胞的细胞质和细胞核中均有存在。然而,Ku70似乎在原肠胚细胞的细胞核中积累的浓度更高。这些结果表明,早期胚胎细胞质中DNA依赖性蛋白激酶活性的逐渐丧失可能是Ku易位至细胞核的结果,这对于合子后转录及随后的细胞分化可能是必要的。