Zhang Suzhen, Cui Xiaoxu, Li Jing, Liang Zhibin, Qiao Wentao, Tan Juan
Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, College of Life Sciences, Nankai University, Tianjin, 300071, China.
Virol Sin. 2016 Apr;31(2):142-9. doi: 10.1007/s12250-015-3652-x. Epub 2016 Mar 11.
Bovine foamy virus (BFV) is a complex retrovirus that infects cattle. Like all retroviruses, BFV encodes a transactivator Tas protein (BTas) that increases gene transcription from viral promoters. BFV encodes two promoters that can interact with BTas, a conserved promoter in the 5' long terminal repeat (LTR) and a unique internal promoter (IP). Our previous study showed that BTas is acetylated by p300 at residues K66, K109, and K110, which markedly enhanced the ability of BTas to bind to DNA. However, whether these residues are important for BFV replication was not determined. Therefore, in this study we provide direct evidence that BTas is required for BFV replication and demonstrate that residues K66, K109, and K110 are critical for BTas function and BFV replication. Full-length infectious clones were generated, which were BTas deficient or contained lysine to arginine (K→R) mutations at position 66, 109, and/or 110. In vivo data indicated that K→R mutations at positions 66, 109, and 110 in BTas impaired transactivation of both the LTR and IP promoters. In addition, the K→R mutations in full-length infectious clones reduced expression of viral proteins, and the triple mutant and BTas deletion completely abrogated viral replication. Taken together, these results indicate that lysine residues at positions 66, 109, and 110 in the BTas protein are crucial for BFV replication and suggest a potential role for BTas acetylation in regulating the viral life cycle.
牛泡沫病毒(BFV)是一种感染牛的复杂逆转录病毒。与所有逆转录病毒一样,BFV编码一种反式激活蛋白Tas(BTas),它可增加病毒启动子的基因转录。BFV编码两个可与BTas相互作用的启动子,一个位于5'长末端重复序列(LTR)中的保守启动子和一个独特的内部启动子(IP)。我们之前的研究表明,BTas在K66、K109和K110位点被p300乙酰化,这显著增强了BTas与DNA结合的能力。然而,这些位点对于BFV复制是否重要尚未确定。因此,在本研究中,我们提供了直接证据表明BTas是BFV复制所必需的,并证明K66、K109和K110位点对于BTas功能和BFV复制至关重要。构建了全长感染性克隆,这些克隆要么缺乏BTas,要么在第66、109和/或110位含有赖氨酸到精氨酸(K→R)的突变。体内数据表明,BTas中第66、109和110位的K→R突变损害了LTR和IP启动子的反式激活。此外,全长感染性克隆中的K→R突变降低了病毒蛋白的表达,三突变体和BTas缺失完全消除了病毒复制。综上所述,这些结果表明BTas蛋白中第66、109和110位的赖氨酸残基对于BFV复制至关重要,并提示BTas乙酰化在调节病毒生命周期中可能发挥作用。