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鸡胚DNA聚合酶γ。从细胞核和线粒体中纯化的γ-聚合酶的同一性。

Chick-embryo DNA polymerase gamma. Identity of gamma-polymerases purified from nuclei and mitochondria.

作者信息

Bertazzoni U, Scovassi A I, Brun G M

出版信息

Eur J Biochem. 1977 Dec 1;81(2):237-48. doi: 10.1111/j.1432-1033.1977.tb11945.x.

Abstract

The level of DNA polymerase gamma as compared to DNA polymerases alpha and beta has been determined in chick embryo by means of specific tests: the amount of gamma-polymerase in the 12-day-old chick embryo reaches about 15% of the total polymerase activity. This enzyme is mainly localized in nuclei and mitochondria, where it represents the prevailing if not the unique DNA polymerase activity. The mitochondrial DNA polymerase gamma is likely to be associated with the internal membrane or the matrix of this organelle since it is not removed by digitonin treatment. The gamma-polymerases have been purified from chick embryo nuclei and mitochondria 500-700 times by means of DEAE-cellulose, phosphocellulose and hydroxyapatite chromatographies. The purified mitochondrial DNA polymerase gamma is closely related to the homologous enzyme purified from the nuclei of the same cells. So far, they cannot be distinguished on the basis of their sedimentation, catalytical properties and response to inhibitors or denaturating agents. The purified gamma enzymes are distinct from the chick embryo DNA polymerases alpha and beta and are not inhibited by antibodies prepared against the latter enzymes. The nuclear and mitochondrial gamma-polymerases do not respond to the oncogenic RNA virus DNA polymerase assay with natural mRNAs.

摘要

通过特定检测方法测定了鸡胚中DNA聚合酶γ与DNA聚合酶α和β相比的水平:12日龄鸡胚中γ聚合酶的量达到总聚合酶活性的约15%。这种酶主要定位于细胞核和线粒体中,在那里它即使不是唯一的也是主要的DNA聚合酶活性。线粒体DNA聚合酶γ可能与该细胞器的内膜或基质相关,因为它不会被洋地黄皂苷处理去除。通过DEAE - 纤维素、磷酸纤维素和羟基磷灰石色谱法,已将鸡胚细胞核和线粒体中的γ聚合酶纯化了500 - 700倍。纯化的线粒体DNA聚合酶γ与从相同细胞的细胞核中纯化的同源酶密切相关。到目前为止,根据它们的沉降、催化特性以及对抑制剂或变性剂的反应,无法区分它们。纯化的γ酶与鸡胚DNA聚合酶α和β不同,并且不受针对后两种酶制备的抗体的抑制。细胞核和线粒体的γ聚合酶对用天然mRNA进行的致癌RNA病毒DNA聚合酶测定没有反应。

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