Facultad de Medicina, Departamento de Microbiología y Parasitología, Universidad Nacional Autónoma de México, Ciudad de México, México.
Departamento de Química, Universidad Autónoma Metropolitana-Iztapalapa, Ciudad de México, México.
PLoS One. 2024 Aug 29;19(8):e0306633. doi: 10.1371/journal.pone.0306633. eCollection 2024.
Transcription regulation in cestodes has been little studied. Here, we characterize the Taenia solium TATA-binding protein (TBP) gene. We found binding sites for transcription factors such as NF1, YY1, and AP-1 in the proximal promoter. We also identified two TATA-like elements in the promoter; however, neither could bind TBP. Additionally, we mapped the transcription start site (A+1) within an initiator and identified a putative downstream promoter element (DPE) located at +27 bp relative to the transcription start site. These two elements are important and functional for gene expression. Moreover, we identified the genes encoding T. solium TBP-Associated Factor 6 (TsTAF6) and 9 (TsTAF9). A Western blot assay revealed that both factors are expressed in the parasite; electrophoretic mobility shift assays and super-shift assays revealed interactions between the DPE probe and TsTAF6-TsTAF9. Finally, we used molecular dynamics simulations to formulate an interaction model among TsTAF6, TsTAF9, and the DPE probe; we stabilized the model with interactions between the histone fold domain pair in TAFs and several pairs of nucleotides in the DPE probe. We discuss novel and interesting features of the TsTAF6-TsTAF9 complex for interaction with DPE on T. solium promoters.
在绦虫中,转录调控的研究较少。在这里,我们对猪带绦虫 TATA 结合蛋白(TBP)基因进行了特征描述。我们在近端启动子中发现了转录因子如 NF1、YY1 和 AP-1 的结合位点。我们还在启动子中鉴定了两个 TATA 样元件;然而,它们都不能与 TBP 结合。此外,我们确定了转录起始位点(A+1)位于起始子内,并鉴定了一个位于转录起始位点后 27 个碱基的推定下游启动子元件(DPE)。这两个元件对于基因表达非常重要和有功能。此外,我们鉴定了编码猪带绦虫 TBP 相关因子 6(TsTAF6)和 9(TsTAF9)的基因。Western blot 分析表明,这两个因子在寄生虫中均有表达;电泳迁移率变动分析和超级迁移率变动分析显示 DPE 探针与 TsTAF6-TsTAF9 之间存在相互作用。最后,我们使用分子动力学模拟构建了 TsTAF6、TsTAF9 和 DPE 探针之间的相互作用模型;我们通过 TAFs 中组蛋白折叠结构域对与 DPE 探针中几个碱基对之间的相互作用来稳定模型。我们讨论了 TsTAF6-TsTAF9 复合物与 T. solium 启动子上 DPE 相互作用的新颖有趣特征。