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Proc Natl Acad Sci U S A. 1985 Apr;82(8):2220-4. doi: 10.1073/pnas.82.8.2220.
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本文引用的文献

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Protein measurement with the Folin phenol reagent.使用福林酚试剂进行蛋白质测定。
J Biol Chem. 1951 Nov;193(1):265-75.
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STUDIES OF THE LIFE CYCLE OF MAMMALIAN CELLS.哺乳动物细胞生命周期的研究。
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Enzymatic synthesis of deoxyribonucleic acid. IX. The polymerase formed after T2 bacteriophage infection of Escherichia coli: a new enzyme.脱氧核糖核酸的酶促合成。IX. T2噬菌体感染大肠杆菌后形成的聚合酶：一种新酶。
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A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid.用于比色法测定脱氧核糖核酸的二苯胺反应的条件及机制研究。
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Purification and partial characterization of a DNA polymerase alpha species from calf thymus.从小牛胸腺中纯化并部分鉴定一种DNA聚合酶α。
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De novo DNA synthesis by a novel mouse DNA polymerase associated with primase activity.一种与引发酶活性相关的新型小鼠DNA聚合酶进行的从头DNA合成。
Nature. 1982 Jul 8;298(5870):180-2. doi: 10.1038/298180a0.
7
Evidence that a high molecular weight replicative DNA polymerase is conserved during evolution.有证据表明，一种高分子量复制性DNA聚合酶在进化过程中是保守的。
Proc Natl Acad Sci U S A. 1981 Nov;78(11):6771-5. doi: 10.1073/pnas.78.11.6771.
8
DNA polymerase alpha mutants from a Drosophila melanogaster cell line.来自黑腹果蝇细胞系的DNA聚合酶α突变体。
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9
Fidelity of mammalian DNA polymerases.哺乳动物DNA聚合酶的保真度。
Science. 1981 Aug 14;213(4509):765-7. doi: 10.1126/science.6454965.
10
Identification of DNA polymerase(s) involved in the repair of viral and cellular DNA in herpes simplex virus type 2-infected cells.鉴定参与2型单纯疱疹病毒感染细胞中病毒和细胞DNA修复的DNA聚合酶。
Virology. 1983 Sep;129(2):524-8. doi: 10.1016/0042-6822(83)90195-2.

海拉细胞DNA聚合酶系统的细胞周期依赖性及特性

Cell-cycle dependence and properties of the HeLa cell DNA polymerase system.

作者信息

Delfini C, Alfani E, De Venezia V, Oberholtzer G, Tomasello C, Eremenko T, Volpe P

出版信息

Proc Natl Acad Sci U S A. 1985 Apr;82(8):2220-4. doi: 10.1073/pnas.82.8.2220.

DOI:10.1073/pnas.82.8.2220
PMID:3857575
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC397528/
Abstract

Analysis of the properties of the DNA polymerase (pol) system as a function of fundamental factors of the assay environment allowed a rather accurate estimation of its dependence on the HeLa cell cycle. For pol alpha, the temperature and pH optima were 38.1 degrees C and 8.0, respectively; for pol beta, these optima were 36.2 degrees C and pH 7.4. Pol gamma showed a pH optimum at 7.7. Optimum activity for both the alpha and beta enzymes was observed at 60 mM Tris. The maximal activity at 36.2 degrees C and pH 7.4 was associated with resistance to N-ethylmaleimide (MalNEt), whereas that at 38.1 degrees C and pH 8.0 was sensitive to MalNEt. Incorporation of [3H]dTTP was maximal after 1 hr of incubation for the former activity and after 4 hr, for the latter. In extracts from cells in early S phase, the pol activity decreased after 1 hr of incubation, was MalNEt-resistant, and was characterized by temperature and pH optima at 36.2 degrees C and 7.4, respectively. In extracts of late S-phase cells, the pol-catalyzed incorporation of [3H]dTTP continued after 4 hr of incubation, was MalNEt-sensitive, and was characterized by temperature and pH optima at 38.1 degrees C and 8.0, respectively. Thus, a pol beta-type activity appeared in early S phase, whereas a pol alpha-type activity appeared in late S. During the G1, M, and G2 phases, a background level of pol activity was observed that showed intermediate kinetic properties.

摘要

通过分析DNA聚合酶（pol）系统的特性与检测环境基本因素之间的关系，可以较为准确地评估其对HeLa细胞周期的依赖性。对于polα，温度和pH的最适值分别为38.1℃和8.0；对于polβ，这些最适值为36.2℃和pH 7.4。Polγ的pH最适值为7.7。在60 mM Tris中观察到α和β酶的最佳活性。在36.2℃和pH 7.4时的最大活性与对N-乙基马来酰亚胺（MalNEt）的抗性相关，而在38.1℃和pH 8.0时的最大活性对MalNEt敏感。对于前一种活性，孵育1小时后[3H]dTTP的掺入量最大，对于后一种活性，孵育4小时后最大。在早S期细胞的提取物中，孵育1小时后pol活性下降，对MalNEt有抗性，其温度和pH最适值分别为36.2℃和7.4。在晚S期细胞的提取物中，孵育4小时后pol催化的[3H]dTTP掺入仍在继续，对MalNEt敏感，其温度和pH最适值分别为38.1℃和8.0。因此，在早S期出现polβ型活性，而在晚S期出现polα型活性。在G1、M和G2期，观察到pol活性的背景水平，其表现出中间动力学特性。