The observation on function and structure of Saccharomyces cerevisiaes Spt5 associates with RNA polymeraseⅡ
| Autor: | Chien-Hung Chung, 鍾建弘 |
|---|---|
| Rok vydání: | 2019 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 107 In the eukaryotes, RNA polymerase Ⅱ is a large protein complex consist of 12 subunits·It catalyzes the transcription of DNA to pre-mRNA、Small nuclear RNA and microRNA. Transcription Factor Spt5 play roles in RNA polymerase Ⅱ mediated transcription, from initiation to termination. Here, we use Saccharomyces cerevisiae system to overexpress galactose inducible Spt5, and characterized its function and interaction with RNA polymerase Ⅱ in a in vitro transcription system. We use Cryo-EM and 3D-reconstruction to demonstrate structure of Spt5/ RNA polymerase Ⅱ complex. Surprisingly, our structure image rollout Spt5 binding with RNA polymerase Ⅱ will lead stalk-like Rpb4/7 heterodimer dissociate from RNA polymerase Ⅱ, and this event will occur during in vivo transcription. In summary, we successfully overexpress functional protein Spt5.Spt5 bind to RNA polymerase Ⅱ will lead Rpb4/7 dissociate from RNA polymerase Ⅱ, this result may reveal a new mechanism of Spt5 to regulate transcription process 。 |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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