Modulation of DNA polymerases α, δ and ε by lactate dehydrogenase and 3-phosphoglycerate kinase
Modulation of DNA polymerases α, δ and ε by lactate dehydrogenase and 3-phosphoglycerate kinase
复制标题
DOI：
10.1016/s0167-4781(97)00229-7
复制
发表时间：
1998-04-01
期刊：
影响因子：
--
通讯作者：
Thielmann, HW
中科院分区：
文献类型：
--
作者：
Popanda, O;Fox, G;Thielmann, HW
有文献报道糖酵解酶（乳酸脱氢酶和3-磷酸甘油酸激酶）存在于哺乳动物细胞核中，在DNA复制、转录和DNA修复中发挥作用。除了它们在细胞质中作为催化剂的作用外。糖酵解酶向细胞核的转移需要修饰，例如磷酸化。我们研究了磷酸化乳酸脱氢酶和3-磷酸甘油激酶对(i)紫外线诱导的DNA修复的影响，使用渗透化的人成纤维细胞，以及（ii）纯化的DNA聚合酶α， δ和epsilon催化增殖大鼠肝脏的体外DNA合成。(i)磷酸化乳酸脱氢酶以剂量依赖的方式刺激正常成纤维细胞中紫外线诱导的DNA修复合成；未磷酸化酶略有抑制。无论乳酸脱氢酶是否被磷酸化，在修复缺陷性着色性干皮病成纤维细胞中，修复性合成都没有增强，这表明修复性DNA合成必须在老年人中可能被刺激。（ii）根据乳酸脱氢酶的磷酸化状态，纯化DNA聚合酶α、δ和epsilon的活性受到不同程度的刺激或抑制。DNA聚合酶也由3-磷酸甘油酸激酶调节，这取决于所使用的引物模板是间隙DNA（模拟DNA合成的修复模式）或单链M13 DNA（代表DNA合成的复制模式）。由于细胞核中的糖酵解酶保留了与其辅因子、细胞质底物和抑制剂的结合能力，因此细胞的能量状态与其复制和修复功能之间可能存在调节联系。(C) 1998爱思唯尔科学有限公司
Literature documents that glycolytic enzymes (among them lactate dehydrogenase and 3-phosphoglycerate kinase) can reside in nuclei of mammalian cells and exert functions in DNA replication, transcription and DNA repair. in addition to their role as catalysts in the cytoplasm. Transfer of glycolytic enzymes to cell nuclei requires modification, for example phosphorylation. We studied the effects of phosphorylated lactate dehydrogenase and 3-phosphoglycerate kinase on (i) UV-induced DNA repair, using permeabilized human fibroblasts, and (ii) in vitro DNA synthesis catalyzed by purified DNA polymerases alpha, delta, and epsilon from proliferating rat liver. (i) Phosphorylated lactate dehydrogenase stimulated UV-induced DNA repair synthesis in normal fibroblasts in a dose-dependent manner; the unphosphorylated enzyme slightly inhibited. In repair-deficient xeroderma pigmentosum fibroblasts reparative synthesis was not enhanced whether lactate dehydrogenase was phosphorylated or not, indicating that reparative DNA synthesis must be possible in older to be stimulated. (ii) Activity of purified DNA polymerases alpha, delta, and epsilon was differentially stimulated or inhibited, according to the phosphorylation status of lactate dehydrogenase. DNA polymerases were also modulated by 3-phosphoglycerate kinase, depending on the primer-templates used which were gapped DNA (mimicking a repair mode of DNA synthesis) or single-stranded M13 DNA (representing the replicative mode of DNA synthesis). Since glycolytic enzymes in cell nuclei retain binding ability for their cofactors, cytoplasmic substrates and inhibitors, a regulatory linkage might exist between the energy state of a cell and its replicative and reparative functions. (C) 1998 Elsevier Science B.V.