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小麦胚萌发过程中DNA合成相关酶和因子的变化

Changes of enzymes and factors involved in DNA synthesis during wheat embryo germination.

作者信息

Benedetto J P, Ech-Chaoui R, Plissonneau J, Laquel P, Litvak S, Castroviejo M

机构信息

Institut de Biochimie et de Génétique Cellulaires, C.N.R.S. 1, Bordeaux, France.

出版信息

Plant Mol Biol. 1996 Sep;31(6):1217-25. doi: 10.1007/BF00040838.

Abstract

We have previously purified and characterized wheat germ DNA polymerases A and B. To determine the role played by DNA polymerases A and B in DNA replication, we have measured the level of their activities during wheat embryo germination. The level of cellular proteins known to be associated with DNA synthesis such as PCNA and DNA primase were also investigated. The activity of DNA polymerase A gradually increased reaching a maximal level at 12 h after germination. Three days later, only a residual activity was detected. DNA polymerase B showed the same pattern during germination with very similar changes in activity. Our results indicate a striking correlation between maximal activities of DNA polymerase A, DNA polymerase B and optimal levels of DNA synthesis. These results support a replicative role of these enzymes. The activity of wheat DNA primase that copurifies with DNA polymerase A also increases during wheat germination. Taking together all its properties, and in spite of its behaviour with some inhibitors. DNA polymerase A may be considered as the plant counterpart of animal DNA polymerase alpha. Concerning DNA polymerase B we have previously shown that PCNA stimulates its processivity. Besides studying the changes of DNA polymerases A and B and DNA primase we have also studied changes in PCNA during germination. We show that PCNA is present in wheat embryos at a constant relatively high level during the first 24 h of germination. After 48 h, the absence of PCNA is concomitant with an important decrease in DNA polymerase B activity. In this report we confirm the behaviour of DNA polymerase B as a delta-like activity.

摘要

我们之前已经纯化并鉴定了小麦胚DNA聚合酶A和B。为了确定DNA聚合酶A和B在DNA复制中所起的作用,我们测定了它们在小麦胚萌发过程中的活性水平。我们还研究了已知与DNA合成相关的细胞蛋白水平,如增殖细胞核抗原(PCNA)和DNA引发酶。DNA聚合酶A的活性在萌发后12小时逐渐增加并达到最高水平。三天后,仅检测到残余活性。DNA聚合酶B在萌发过程中表现出相同的模式,活性变化非常相似。我们的结果表明,DNA聚合酶A、DNA聚合酶B的最大活性与DNA合成的最佳水平之间存在显著相关性。这些结果支持了这些酶的复制作用。与DNA聚合酶A共纯化的小麦DNA引发酶的活性在小麦萌发过程中也增加。综合其所有特性,尽管它对某些抑制剂有反应,但DNA聚合酶A可被视为动物DNA聚合酶α的植物对应物。关于DNA聚合酶B,我们之前已经表明PCNA可刺激其持续合成能力。除了研究DNA聚合酶A和B以及DNA引发酶的变化外,我们还研究了萌发过程中PCNA的变化。我们发现,在萌发的前24小时内,PCNA在小麦胚中以相对较高的恒定水平存在。48小时后,PCNA的缺失与DNA聚合酶B活性的显著下降同时出现。在本报告中,我们证实了DNA聚合酶B作为一种类似δ活性的行为。

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