- 作者: Gesa Jonas Silke Hoffmann Dieter Willbold
- 作者服務機構: Institut fur molekulare biotechnologie, jena, Universitat bayreuth, lehrstuhl fur biopolymere, Bayreuth, Deutschland
- 中文摘要: --
- 英文摘要: The transactivator protein (Tat) of the human immuno-deficiency virus (HIV) is a key regulatory protein in theviral replication cycle. Together with cellular cyclin T1and an RNA element (transactivation response; TAR)located at the 5' end of all viral transcripts, it forms a ter-nary complex that ultimately enhances the expression ofall viral genes. In this ternary complex, cyclin T1 interactsdirectly with Tat and TAR. The presence of cyclin T1 isessential for high TAR RNA affinity and specificity of Tat.To study protein-protein and protein-RNA interaction,we developed a phage display system that displays func-tional Tat on the surface of bacteriophage M13. The addi-tion of recombinant cyclin T1 to the selections yielded aphage display system that mirrors all binding propertiesof the cyclin T1-Tat-TAR complex known from cell assaysand biochemical studies. Phage-displayed Tat protein aswell as the cyclin T1 are fully functional. The relativebinding capabilities of wild-type- and mutant Tat-dis-playing phages show that the presence of cyclin T1 sig-nificantly reduces the importance of basic residues in thebasic sequence region of Tat for its binding to TAR.
- 中文關鍵字: --
- 英文關鍵字: HIV-1 Transactivator. Cyclin T1. Phage Display. Protein-RNA interactions