Two promoters, LTR (long terminal repeat) and IP (internal promoter), exist in genomes of foamy viruses. Cell transfection and transient expression assay demonstrated that: the basal activity of BFV (bovine foamy viru...Two promoters, LTR (long terminal repeat) and IP (internal promoter), exist in genomes of foamy viruses. Cell transfection and transient expression assay demonstrated that: the basal activity of BFV (bovine foamy virus) IP is much higher than that of BFV LTR; transactivator of BFV—Borf 1, which activates gene expression directed by BFV LTR, also functions on BFV IP with an activation fold higher than that on LTR. The results suggest that BFV IP and LTR may regulate viral gene expression by different mechanisms, and that Borf 1 may stimulate BFV IP and LTR in different ways. In addition, an in vivo DNA competition assay demonstrated that a common transcription factor may be involved in both mechanisms of the two promoters.展开更多
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 promote...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.展开更多
文摘Two promoters, LTR (long terminal repeat) and IP (internal promoter), exist in genomes of foamy viruses. Cell transfection and transient expression assay demonstrated that: the basal activity of BFV (bovine foamy virus) IP is much higher than that of BFV LTR; transactivator of BFV—Borf 1, which activates gene expression directed by BFV LTR, also functions on BFV IP with an activation fold higher than that on LTR. The results suggest that BFV IP and LTR may regulate viral gene expression by different mechanisms, and that Borf 1 may stimulate BFV IP and LTR in different ways. In addition, an in vivo DNA competition assay demonstrated that a common transcription factor may be involved in both mechanisms of the two promoters.
基金supported by grants from the National Natural Science Foundation of China(Na.31370182)Tianjin Research Program and Application Foundation and Advanced Technology(Na.12YFQNJC02300)111 Project(B08011)
文摘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.
