Eukaryotic RNA Polymerase III Transcription

We are interested in understanding the molecular mechanisms of eukaryotic RNA polymerase (pol) transcription. The current focus is on the pol III transcription system that synthesizes transfer RNAs (tRNA) and many other small structural RNAs such as 5S rRNA and U6 spliceosomal RNA. We have employed multiple site-directed mutational and biochemical approaches to study the protein subunits of pol III and transcription factors TFIIIB and TFIIIC from the budding yeast, Saccharomyces cerevisiae. In particular, we apply a site-specific photo-cross-linking method to map protein interactions within the protein-nucleic acid complexes during the transcription process. Our studies have revealed specific structural regions in the Rpc53, Rpc37, Rpc82, and Rpc31 subunits of pol III that are involved in transient protein interactions for the initiation, elongation, and termination stages of the transcription cycle. Our research thus provides functional and structural insights into the macromolecular assemblies of RNA polymerase transcription.

• 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, Academia Sinica Career Development Award

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.