Arnosti D N, Preston C M, Hagmann M, Schaffner W, Hope R G, Laughlan G, Luisi B F
Institut für Molekularbiologie II, Universitat Zürich, Switzerland.
Nucleic Acids Res. 1993 Dec 11;21(24):5570-6. doi: 10.1093/nar/21.24.5570.
The herpes simplex virus protein VP16 interacts with cellular factors, including the protein Oct-1, to activate viral immediate early (IE) gene transcription. We have reproduced this effect by addition of purified, full-length VP16 and the DNA-binding 'POU' domain of Oct-1 (Oct-1/POU) to a HeLa cell in vitro transcription system. Stimulation of transcription was dependent on the IE-specific element, TAATGARAT. In agreement with earlier observations from electrophoretic mobility shift assays, activation was not observed when Oct-2/POU, the DNA-binding domain from the Oct-2 protein, was substituted for Oct-1/POU. Single round transcription assays revealed that, together, VP16 and Oct-1/POU facilitate the assembly of pre-initiation complexes at target gene promoters.
单纯疱疹病毒蛋白VP16与包括Oct-1蛋白在内的细胞因子相互作用,以激活病毒即刻早期(IE)基因转录。我们通过向体外HeLa细胞转录系统中添加纯化的全长VP16和Oct-1的DNA结合“POU”结构域(Oct-1/POU)重现了这种效应。转录的刺激依赖于IE特异性元件TAATGARAT。与早期电泳迁移率变动分析的观察结果一致,当用Oct-2蛋白的DNA结合结构域Oct-2/POU替代Oct-1/POU时,未观察到激活。单轮转录分析表明,VP16和Oct-1/POU共同促进了靶基因启动子处起始前复合物的组装。