Ikeda J E, Enomoto T, Hurwitz J
Proc Natl Acad Sci U S A. 1981 Feb;78(2):884-8. doi: 10.1073/pnas.78.2.884.
A protein fraction isolated from the cytosol of adenovirus-infected HeLa cells, which contained DNA polymerase alpha, catalyzed adenoviral DNA replication in the presence of adenovirus DNA binding protein, eukaryotic DNA polymerase beta, ATP, all four dNTPs, and MgCl2. DNA replication started at either end of exogenously added adenoviral DNA and was totally dependent on the presence of terminal 55,000-dalton proteins on the DNA template. The replicaton of adenovirus DNA in the system was sensitive to aphidicolin and retained nearly all the properties of DNA replication that occur in vivo or in vitro with crude extracts. The 5'-ends of the newly synthesized adenovirus DNA strands were covalently linked to an 80,000-dalton protein linked to dCMP. DNA synthesized with purified proteins was only 25-50+ the length of parental viral strands. Addition of cytosol extracts from uninfected HeLa cells to reaction mixtures containing purified proteins gave full-length adenoviral DNA strands.
从腺病毒感染的HeLa细胞胞质溶胶中分离出的一种蛋白质组分，其含有DNA聚合酶α，在腺病毒DNA结合蛋白、真核DNA聚合酶β、ATP、所有四种脱氧核苷三磷酸和氯化镁存在的情况下催化腺病毒DNA复制。DNA复制在外源添加的腺病毒DNA的任一端开始，并且完全依赖于DNA模板上末端55,000道尔顿蛋白质的存在。该系统中腺病毒DNA的复制对阿非科林敏感，并保留了体内或体外粗提物中发生的DNA复制的几乎所有特性。新合成的腺病毒DNA链的5′末端与连接有dCMP的80,000道尔顿蛋白质共价连接。用纯化蛋白质合成的DNA仅为亲本病毒链长度的25 - 50倍。向含有纯化蛋白质的反应混合物中添加未感染HeLa细胞的胞质溶胶提取物可得到全长腺病毒DNA链。
