Avidan Orna, Meer Michal Entin, Oz Iris, Hizi Amnon
Department of Cell Biology and Histology, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
Eur J Biochem. 2002 Feb;269(3):859-67. doi: 10.1046/j.0014-2956.2001.02719.x.
We have recently expressed in bacteria the enzymatically active reverse transcriptase (RT) of bovine leukemia virus (BLV) [Perach, M. & Hizi, A. (1999) Virology 259, 176-189]. In the present study, we have studied in vitro two features of the DNA polymerase activity of BLV RT, the processivity of DNA synthesis and the fidelity of DNA synthesis. These properties were compared with those of the well-studied RTs of human immunodeficiency virus type 1 (HIV-1) and murine leukaemia virus (MLV). Both the elongation of the DNA template and the processivity of DNA synthesis exhibited by BLV RT are impaired relative to the other two RTs studied. Two parameters of fidelity were studied, the capacity to incorporate incorrect nucleotides at the 3' end of the nascent DNA strand and the ability to extend these 3' end mispairs. BLV RT shows a fidelity of misinsertion higher than that of HIV-1 RT and lower than that of MLV RT. The pattern of mispair elongation by BLV RT suggests that the in vitro error proneness of BLV RT is closer to that of HIV-1 RT. These fidelity properties are discussed in the context of the various retroviral RTs studied so far.
我们最近已在细菌中表达了牛白血病病毒(BLV)具有酶活性的逆转录酶(RT)[佩拉赫,M. & 希齐,A.(1999年)《病毒学》259,176 - 189]。在本研究中,我们在体外研究了BLV RT的DNA聚合酶活性的两个特征,即DNA合成的持续性和DNA合成的保真度。将这些特性与研究充分的1型人类免疫缺陷病毒(HIV - 1)和鼠白血病病毒(MLV)的RTs特性进行了比较。相对于所研究的其他两种RTs,BLV RT所表现出的DNA模板延伸和DNA合成的持续性均受到损害。研究了保真度的两个参数,即在新生DNA链3'端掺入错误核苷酸的能力以及延伸这些3'端错配的能力。BLV RT显示出的错配插入保真度高于HIV - 1 RT且低于MLV RT。BLV RT错配延伸的模式表明,BLV RT在体外的易出错性更接近HIV - 1 RT。在迄今所研究的各种逆转录病毒RTs的背景下讨论了这些保真度特性。