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真核生物中 POPs、植物细胞器 DNA 聚合酶的保守性。

Conservation of POPs, the plant organellar DNA polymerases, in eukaryotes.

机构信息

Department of Plant Sciences, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan; Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan.

出版信息

Protist. 2011 Jan;162(1):177-87. doi: 10.1016/j.protis.2010.06.001. Epub 2010 Jul 21.

Abstract

POPs, plant organellar DNA polymerases, have been isolated from various photosynthetic eukaryotes. Previously, we purified the native POP of Cyanidioschyzon merolae(CmPOP) from whole cellular extracts and showed that CmPOP has DNA polymerase activity with a high processivity and a 3'-5' exonuclease activity, and its expression is related to cell proliferation. In rice, the recombinant protein of POP has activities found in CmPOP, and high fidelity of POP has also been demonstrated. These facts suggest that POPs are involved in the replication of organellar genomes. POPs are also conserved in most non-opisthokont eukaryotes, which lack DNA polymerase γ (Polγ), a mitochondrial replication enzyme in opisthokonts (fungi and animals). The ciliateTetrahymena thermophilacontains a single gene for a putative POP (TetPOP). Immunoblot analysis demonstrated that TetPOP is localized in mitochondria, and TetPOP has been purified from mitochondria through a column chromatography series. Sensitivity to phosphonoacetate and dideoxyTTP was examined in POPs (TetPOP and CmPOP) or POP-containing organelles (chloroplasts of Arabidopsis) and other polymerases (DNA polymerase I and mitochondria of rat liver, which contain Polγ), and the results suggest that high sensitivity to phosphonoacetate is unique to POPs in Family-A DNA polymerases. Finally, we propose a model for the succession of organellar DNA polymerases.

摘要

POPs,即植物细胞器 DNA 聚合酶,已从各种光合真核生物中分离出来。此前,我们从全细胞提取物中纯化了嗜热四膜虫(Cyanidioschyzon merolae)的天然 POP(CmPOP),并证明 CmPOP 具有高延伸性和 3'-5'外切核酸酶活性的 DNA 聚合酶活性,其表达与细胞增殖有关。在水稻中,POP 的重组蛋白具有在 CmPOP 中发现的活性,并且已经证明了 POP 的高保真度。这些事实表明,POPs 参与了细胞器基因组的复制。POPs 在大多数非后口动物真核生物中也保守存在,而后口动物(真菌和动物)具有线粒体复制酶 DNA 聚合酶 γ(Polγ)。纤毛虫嗜热四膜虫含有一个单一的假定 POP(TetPOP)基因。免疫印迹分析表明 TetPOP 定位于线粒体中,并且已经通过一系列柱层析从线粒体中纯化了 TetPOP。在 POP(TetPOP 和 CmPOP)或含有 POP 的细胞器(拟南芥的叶绿体)和其他聚合酶(含有 Polγ 的 DNA 聚合酶 I 和大鼠肝脏的线粒体)中检查了对膦酸乙酸和双脱氧 TTP 的敏感性,结果表明对膦酸乙酸的高敏感性是 A 家族 DNA 聚合酶中 POP 的独特特征。最后,我们提出了一个细胞器 DNA 聚合酶的接替模型。

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