12型人腺病毒pTP-dCMP起始复合物的体外形成

Formation in vitro of the pTP-dCMP initiation complex of human adenovirus type 12.

作者信息

Chowrira B M, Zhao J S, Lucher L A

机构信息

Department of Biological Sciences, Illinois State University, Normal 61761.

出版信息

J Gen Virol. 1991 Feb;72 ( Pt 2):427-30. doi: 10.1099/0022-1317-72-2-427.

DOI:10.1099/0022-1317-72-2-427

PMID:1993880

Abstract

We report the covalent addition of [32P]dCMP to a protein from group A adenovirus 12 (Ad12)-infected human (KB) cells in vitro, using crude extracts. Synthesis of the 60K protein-dCMP complex required a DNA template containing a terminally located adenovirus replication origin; the protein-dCMP bond was alkali-labile but acid-stable. We therefore conclude that this product is the Ad12 terminal protein precursor (pTP)-dCMP initiation complex for DNA replication. Synthesis of Ad12 pTP-dCMP was specific for dCTP but was stimulated by dATP. In contrast to Ad2, the Ad12 initiation reaction required ATP. Antipeptide antiserum targeted to Ad DNA polymerase inhibited Ad12 pTP-dCMP synthesis in vitro, providing evidence that Ad DNA polymerase catalyse dCMP addition to pTP during initiation.

摘要

我们报道了在体外使用粗提物将[32P]dCMP共价添加到感染A组腺病毒12型（Ad12）的人（KB）细胞中的一种蛋白质上。60K蛋白质-dCMP复合物的合成需要一个含有位于末端的腺病毒复制起点的DNA模板；蛋白质-dCMP键对碱不稳定但对酸稳定。因此，我们得出结论，该产物是用于DNA复制的Ad12末端蛋白前体（pTP）-dCMP起始复合物。Ad12 pTP-dCMP的合成对dCTP具有特异性，但受到dATP的刺激。与Ad2不同，Ad12起始反应需要ATP。靶向Ad DNA聚合酶的抗肽抗血清在体外抑制Ad12 pTP-dCMP的合成，这证明Ad DNA聚合酶在起始过程中催化dCMP添加到pTP上。