Richard Burgess
Professor of Oncology
Ph.D., Harvard University, 1969
Postdoctoral Research: University of Geneva, Switzerland
Address: 407 McArdle
Telephone: 263-2635
E-mail: burgess@oncology.wisc.edu
Research Interests:
E. coli & eukaryotic RNA polymerases, transcription factors, and gene regulation
Research Description: Work in my lab focuses on RNA polymerases (RNAPs, transcription factors and their roles in regulating RNA synthesis. We study the E. coli RNAP sigma70 subunit, by a concerted use of protein and physical chemistry, monoclonal antibodies (MAbs), molecular genetics, computer-based sequence and structure analysis, and biochemistry. We have developed a powerful new methods, using histidine-tagged RNAP subunit fragments and “far-western blotting,” to map interaction domains and have identified a major binding site for sigma70 within the region of amino acids 260-309 of the beta subunit of core RNAP. Site-directed mutagenesis of this region allowed us to identify amino acids residues critical for sigma binding in vitro and in vivo.
Understanding these interactions in detail will allow us to design potentially important new antibiotics that work by disrupting transcription in pathogenic bacteria.
We are utilizing special “polyolresponsive” MAbs that we have discovered to immunoaffinity purify RNAP II from human and yeast cells. We are focusing on the subunit architecture of yeast RNAP II and the interactions of the RNAP subunits with transcription factors and DNA. Using cloned and overproduced yeast RNAP II subunits, we are mapping specific subunit-subunit interactions and trying to reconstitute the active enzyme. We are studying the interactions of human RNAP II with the basal transcription factors, TBP and TFIIB, at a minimal promoter.
Representative Publications:
Lamberski, J.A., Thompson, N.E. and Burgess, R.R. 2006. Expression and purification of a single-chain variable fragment antibody derived from a polyol-responsive monoclonal antibody. Protein Expr Purif. 47:82-92.
Raffaelle, M., Kanin, E.I., Vogt, J., Burgess, R.R. and Ansari, A.Z. 2005. Holoenzyme switching and stochastic release of sigma factors from RNA polymerase in vivo. Mol Cell. 20:357-66.
Kozlov, M., Bergendahl, V., Burgess, R., Goldfarb, A. and Mustaev, A. 2005. Homogeneous fluorescent assay for RNA polymerase. Anal Biochem. 342:206-213.
Bergendahl, V. and Burgess, R.R. 2003. Studying sigma-core interactions in Escherichia coli RNA polymerase by electrophoretic shift assays and luminescence resonance energy transfer. Methods Enzymol. 370:192-205.
Thompson, N.E., Arthur, T.M. and Burgess, R.R. 2003. Development of an epitope tag for the gentle purification of proteins by immunoaffinity chromatography: application to epitope-tagged green fluorescent protein. Anal Biochem. 323:171-179.
Professor of Oncology
Ph.D., Harvard University, 1969
Postdoctoral Research: University of Geneva, Switzerland
Address: 407 McArdle
Telephone: 263-2635
E-mail: burgess@oncology.wisc.edu
Research Interests:
E. coli & eukaryotic RNA polymerases, transcription factors, and gene regulation
Work in my lab focuses on RNA polymerases (RNAPs, transcription factors and their roles in regulating RNA synthesis. We study the E. coli RNAP sigma70 subunit, by a concerted use of protein and physical chemistry, monoclonal antibodies (MAbs), molecular genetics, computer-based sequence and structure analysis, and biochemistry. We have developed a powerful new methods, using histidine-tagged RNAP subunit fragments and “far-western blotting,” to map interaction domains and have identified a major binding site for sigma70 within the region of amino acids 260-309 of the beta subunit of core RNAP. Site-directed mutagenesis of this region allowed us to identify amino acids residues critical for sigma binding in vitro and in vivo.
Lamberski, J.A., Thompson, N.E. and Burgess, R.R. 2006. Expression and purification of a single-chain variable fragment antibody derived from a polyol-responsive monoclonal antibody. Protein Expr Purif. 47:82-92.
