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Volume 23 Issue 1
February 2008

    Juan TAN, Kai WU, Rui CHANG, Qi-min CHEN, Yun-qi GENG and Wen-tao QIAO. Subcellular Localization Analysis of Bovine Foamy Virus Borf1 Protein[J]. Virologica Sinica, 2008, 23(1): 37-42. doi: 10.1007/s12250-008-2893-3
    Citation: Juan TAN, Kai WU, Rui CHANG, Qi-min CHEN, Yun-qi GENG, Wen-tao QIAO. Subcellular Localization Analysis of Bovine Foamy Virus Borf1 Protein .VIROLOGICA SINICA, 2008, 23(1) : 37-42.  http://dx.doi.org/10.1007/s12250-008-2893-3
    shu

    牛泡沫病毒Borf-1蛋白亚细胞定位研究

    cstr: 32224.14.s12250-008-2893-3

    • 南开大学生命科学学院，天津300071

    • Borf1蛋白是牛泡沫病毒（BFV）侵入细胞后早期表达的蛋白，在病毒的生活周期中起着至关重要的作用。Borf-1蛋白为DNA结合蛋白，它通过结合在病毒基因组的两类启动子LTR和IP的相应应答元件上行使其反式激活功能，调控病毒基因表达。为了研究Borf1在病毒感染过程的定位，本文通过克隆、表达并纯化可溶性Borf-1蛋白后，免疫小鼠获得高效价抗血清。通过蛋白印迹实验，与商品化标签抗体比较，结果显示抗血清可以灵敏并特异性识别真核表达的Borf1蛋白。利用获得的多克隆抗体分别首先检测了转染的Borf1蛋白在HeLa细胞中的定位，免疫荧光实验显示Borf1蛋白在HeLa细胞中呈现核浆共分布的状态。然后，进一步检测了BFV感染的胎牛肺细胞（FBL）中Borf1的定位，结果显示在BFV感染的过程中，Borf1蛋白也呈现核浆共分布状态。

    Subcellular Localization Analysis of Bovine Foamy Virus Borf1 Protein

    • The Key Laboratory of Molecular Microbiology and Techaology, Ministry of Education and Tianjin Key Laboratory of Microbial Functional Genomics, College of Life Sciences, Nankai University, Tianjin 300071, China

    • Corresponding author: Wen-tao QIAO, wentaoqiao@nankai.edu.cn
    • Received Date: 23 July 2007
      Accepted Date: 20 December 2007

      Fund Project: National Natural Science Foundation of China 30770097Natural Science Foundation of Tianjin 05YFJM-JC01000National Natural Science Foundation of China 30570072

    • The Borf1 protein is encoded by an immediate-early gene of the bovine foamy virus (BFV) and plays a key role in the viral life cycle. Borf1 is a DNA binding protein which can transactivate both the long terminal repeat (LTR) and the internal promoter (IP) of BFV by specifically binding to the transactivation responsive element (TRE). To analyze the subcellular localization of Borf1 during the BFV life cycle, this gene was cloned into a prokaryotic expression vector and expressed in a soluble form. After the purification and immunization, we raised the mouse anti-Borf1 serum with a high titer based on ELISA results. Western blot analysis showed that the antiserum could specifically recognize the Borf1 protein that was expressed in 293T cells. With this specific serum, we revealed the nuclear and cytoplasmic localization of Borf1 in HeLa cells that was transfected with Borf1. Moreover, the immuno-fluorescence assay also showed that the localization of Borf1 during the infection and transfection of BFV was identical.
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      1. Lecellier C H, Saib A. 2000. Foamy viruses: between retroviruses and pararetroviruses. Virology, 271: 1-8.
          doi: 10.1006/viro.2000.0216

      2. Liu J J, Liu S H, Chen Q M, et al. 1999. Borf-1 protein identified as a transcriptional trans-activator of bovine foamy virus. Chin Sci Bull, 44 (11): 1017-1021. (in Chinese)
          doi: 10.1007/BF02886021

      3. Liu J J, Wang S Z, Zhang L, et al. 2000. Functional Analysis of Bovine Foamy Virus Accessory Protein and Borf1 Responsive Elements in Long Terminal Repeat. Chin J Virol, 16 (2): 141-149. (in Chinese)

      4. Lochelt M. 2003. Foamy virus transactivation and gene expression. Curr Top Microbiol Immunol, 277: 27-61.

      5. Meiering C D, Linial M L. 2002. Reactivation of a complex retrovirus is controlled by a molecular switch and is inhibited by a viral protein. Proc Natl Acad Sci USA, 99 (23): 15130-15135.
          doi: 10.1073/pnas.242491999

      6. Meiering C D, Linial M L. 2003. The promyelocytic leukemia protein does not mediate foamy virus latency in vitro. J Virol, 77 (3): 2207-2213.
          doi: 10.1128/JVI.77.3.2207-2213.2003

      7. Switzer W M, Bhullar V, Shanmugam V, et al. 2004. Frequent Simian Foamy Virus Infection in Persons Occupationally Exposed to Nonhuman Primates. J Virol, 78 (6): 2780-2789.
          doi: 10.1128/JVI.78.6.2780-2789.2004

      8. Zhang L, Qiao W T, Liu S H, et al. 2000. Regulatory Elements That Mediate Transactivation by Tas in the Internal Promoter of Bovine Foamy Virus. Chin J Virol, 16 (3): 232-236. (in Chinese)

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      Subcellular Localization Analysis of Bovine Foamy Virus Borf1 Protein

        Corresponding author: Wen-tao QIAO, wentaoqiao@nankai.edu.cn
      • The Key Laboratory of Molecular Microbiology and Techaology, Ministry of Education and Tianjin Key Laboratory of Microbial Functional Genomics, College of Life Sciences, Nankai University, Tianjin 300071, China
      • Received Date: 23 July 2007
      • Accepted Date: 20 December 2007
      Fund Project:  National Natural Science Foundation of China 30770097Natural Science Foundation of Tianjin 05YFJM-JC01000National Natural Science Foundation of China 30570072

      Abstract: The Borf1 protein is encoded by an immediate-early gene of the bovine foamy virus (BFV) and plays a key role in the viral life cycle. Borf1 is a DNA binding protein which can transactivate both the long terminal repeat (LTR) and the internal promoter (IP) of BFV by specifically binding to the transactivation responsive element (TRE). To analyze the subcellular localization of Borf1 during the BFV life cycle, this gene was cloned into a prokaryotic expression vector and expressed in a soluble form. After the purification and immunization, we raised the mouse anti-Borf1 serum with a high titer based on ELISA results. Western blot analysis showed that the antiserum could specifically recognize the Borf1 protein that was expressed in 293T cells. With this specific serum, we revealed the nuclear and cytoplasmic localization of Borf1 in HeLa cells that was transfected with Borf1. Moreover, the immuno-fluorescence assay also showed that the localization of Borf1 during the infection and transfection of BFV was identical.

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