Eukaryotic RNA Polymerase III Transcription

本實驗室研究主題為真核細胞核醣核酸聚合酶（eukaryotic RNA polymerase）的蛋白質結構與功能，核醣核酸聚合酶由許多次單元蛋白質組成，並且需要結合許多轉錄因子，進行核醣核酸的合成。我們以生物化學方法解出第三型核醣核酸聚合酶（RNA Polymerase III）的蛋白質反應網路，此研究成果，解釋許多蛋白質的重要結構區塊如何調控轉錄起始、延展與終止，此研究成果也促進其他研究團隊解出轉錄起始複合體的結構。 研究成果更被引用於解釋因第三型核醣核酸聚合酶系統的基因異常，所造成的癌症、神經退化、發展遲滯、老化等問題。本實驗室的研究成果，更進而用於比較真核細胞中各轉錄系統的結構與調控機制之異同，對於基因表現、細胞生長、及疾病等生物醫學領域的分子機制研究有深遠影響。本實驗室的也同時發展以單分子生物物理方法，研究核醣核酸聚合酶的動態結構，此計畫為與中研院化學所章為皓實驗室的跨領域合作研究，目前已建立以非自然性胺基酸法聯結螢光染劑於蛋白質，於單分子螢光能量轉移實驗中，觀察到核醣核酸聚合酶結構的動態與改變，此研究可用於觀察蛋白質於轉錄時的結構變化，有助於深入瞭解個別轉錄基因的功能及解釋轉錄的基本機制。

• Education
• Achievements & Honors
• Selected publications
• Lab members

• PDF, 2000-2007, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
• Ph.D., 2000, Chemistry, University of Georgia Georgia, USA
• MS, 1995, Chemistry, University of Texas at El Paso, Texas, USA
• BS, 1989, Chemistry, Soochow University, Taipei, Taiwan

• 2009-2013, 中研院前瞻計畫

1. Wu, J.-S., Chen, T.-Y., Lin, S.-Y., Lin, S.-Y., Hung, C.- Y., Tu, I.-P., Chen, H.-T., Chang, W.-H. (2019) Deriving a sub-nanomolar affinity peptide from TAP to enable smFRET analysis of RNA polymerase II complexes. Methods 159-160: 59-69.
2. Khoo, S.-K., Wu, C.-C., Lin, Y.-C., Chen, H.-T. (2018) The TFIIE-related Rpc82 subunit of RNA polymerase III interacts with the TFIIB-related transcription factor Brf1 and the polymerase cleft for transcription initiation. Nucleic Acids Research 46: 1157-1166.
3. Wei, Y.-Y., Chen, H.-T. (2018) Functions of the TFIIErelated tandem winged-helix domain of Rpc34 in RNA polymerase III initiation and elongation. Molecular and Cellular Biology 38: e00105-17.
4. Hu, H.-L., Wu, C.-C., Lee, J.-C., Chen, H.-T. (2015) A region of Bdp1 necessary for transcription initiation that is located within the RNA Polymerase III active site cleft. Molecular and Cellular Biology 35: 2831-2840.
5. Khoo, S.-K., Wu, C.-C., Lin, Y.-C., Lee, J.-C., Chen, H.-T. (2014) Mapping the protein interaction network for the TFIIB-related factor Brf1 in the RNA polymerase III preinitiation complex. Molecular and Cellular Biology 34: 551-559.
6. Wu, C.-C., Herzog, F., Jennebach, S., Lin, Y.-C., Pai, C.-Y., Aebersold, R., Cramer, P., Chen, H.-T. (2012) RNA polymerase III subunit architecture and implications for open promoter complex formation. PNAS 109: 19232-19237.
7. Wu, C.-C., Lin, Y.-C., Chen, H.-T. (2011) The TFIIF-like Rpc37/53 Dimer Lies at the Center of a Protein Network to Connect TFIIIC, Bdp1, and the RNA Polymerase III Active Center. Molecular and Cellular Biology 31: 2715- 2728.