Tsurumi T, Yamada H, Daikoku T, Yamashita Y, Nishiyama Y
Laboratory of Viral Oncology, Aichi Cancer Center Research Institute, Nagoya, Japan.
Biochem Biophys Res Commun. 1997 Sep 8;238(1):33-8. doi: 10.1006/bbrc.1997.7234.
The Epstein-Barr virus (EBV) DNA polymerase (Pol) holoenzyme is an essential enzyme required for ori-Lyt dependent EBV DNA replication. Using singly primed M13ssDNA circles as template, the EBV DNA Pol holoenzyme synthesized DNA chains greater than the unit length of M13 ssDNA in addition to full length products even at a low ratio of polymerase molecule per templates. The long replication products consisted of circular double-stranded DNA with single-stranded tails that were sensitive to mung bean nuclease. Reconstitution of the EBV Pol holoenzyme by preincubation of BALF5 Pol catalytic subunit and BMRF1 Pol accessory subunit in vitro resulted in reproduction of the strand displacement DNA synthesis. Thus, the EBV DNA Pol holoenzyme by itself is able to produce strand displacement coupled to the polymerization process in a highly processive way in the absence of any other protein.
爱泼斯坦-巴尔病毒(EBV)DNA聚合酶(Pol)全酶是ori-Lyt依赖性EBV DNA复制所必需的一种关键酶。以单引物M13单链DNA环为模板,即使在聚合酶分子与模板的比例很低时,EBV DNA Pol全酶除了能合成全长产物外,还能合成大于M13单链DNA单位长度的DNA链。这些长复制产物由带有对绿豆核酸酶敏感的单链尾巴的环状双链DNA组成。通过在体外预孵育BALF5 Pol催化亚基和BMRF1 Pol辅助亚基来重建EBV Pol全酶,可实现链置换DNA合成的重现。因此,EBV DNA Pol全酶自身能够在没有任何其他蛋白质的情况下,以高度持续的方式产生与聚合过程偶联的链置换。