David Brow
Professor of Biomolecular Chemistry
Ph.D., University of California at San Diego, 1986
Postdoctoral Research: University of California at San Francisco
Address: 4204B Biochemical Sciences
Telephone: 262-1475
E-mail: dabrow@wisc.edu
Research Interests:
Nuclear steps in eukaryotic gene expression
Research Fields:
Gene Expression
Yeast and Fungi
Research Description: We investigate the molecular mechanisms of eukaryotic gene expression, with emphasis on transcription and pre-mRNA splicing. Because the fundamental mechanisms of gene expression are conserved among eukaryotes, we have chosen the genetically tractable yeast S. cerevisiae as a model system. Our transcription studies focus on how RNA polymerases (Pol) II and III identify a gene, and on the detailed mechanism of initiation and termination of transcription by Pol II. We use genetic, genomic, and biochemical approaches in these studies. For example, we have measured Pol II occupancy across the entire yeast genome at 100-200 base-pair resolution by chromatin immunoprecipitation, and we have identified mutations in Pol II that cause it to “read-through” transcription terminators. We discovered and are characterizing a Pol II termination pathway that uses the helicase Sen1 and appears to be most important for short transcripts. In our splicing studies we examine the mechanism and function of dynamic RNA-RNA and RNA-protein interactions in assembling the active site of the spliceosome. We defined a complex network of genetic interactions that involve two RNAs (U4 and U6), two helicases, an RNA-binding protein, and the largest and most conserved spliceosomal protein (Prp8). We use detailed mutational studies, in vitro splicing and RNA-binding assays, and structural biology to dissect the mechanism of spliceosome activation. Mutations in the human Prp8 and Sen1 genes cause retinal degeneration and progressive motor neuron dysfunction, respectively. We hope our studies will illuminate the mechanisms of these diseases.
Representative Publications:
Brow, D.A.
(2011). Sen-sing RNA terminators. Molec. Cell 42, 717-718.
Martin-Tumasz,
S., Richie, A.C., Clos, L.J. II, Brow†, D.A., and Butcher†,
S.E. (2011). A novel occluded RNA
recognition motif in Prp24 unwinds the U6 RNA internal stem loop. Nucl.
Acids Res. doi: 10.1093/nar/gkr455.
†corresponding authors
Brow,
D.A. Eye on RNA unwinding. (2009). Nature Struct. Molec. Biol. 16, 7-8.
Kuehner,
J.N. and Brow, D.A. Regulation of a eukaryotic gene
by GTP-dependent start site selection and transcription attenuation. (2008).
Molec. Cell 31, 201-211. (Cover
article)
McManus,
C.J., Schwartz, M.L., Butcher, S.E. and Brow, D.A. (2007). A dynamic bulge in
the U6 RNA internal stem-loop functions in spliceosome assembly and activation.
RNA 13, 2252-2265.
Bae, E.,
N.J. Reiter, C.A. Bingman, S.S. Kwan, D. Lee, G.N. Phillips Jr., S.E. Butcher,
and D.A. Brow. (2007). Structure and interactions of the first three RNA
recognition motifs of splicing factor Prp24. J. Mol. Biol., 1447-1458.
Steinmetz,
E.J., C.L. Warren, J.N. Kuehner, B. Panbehi, A.Z. Ansari, and D.A. Brow.
(2006). Genome-wide distribution of yeast RNA polymerase II and its control by
Sen1 helicase. Molec. Cell 24,
735-746. (Cover article)
Kuehner,
J.N, and D.A. Brow. (2006). Quantitative analysis of in vivo initiator selection by yeast RNA polymerase II supports a
scanning model. J. Biol. Chem. 281,
14119-14128.
Steinmetz,
E.J., S.B.H. Ng, J.P. Cloute, and D.A. Brow. (2006). Cis- and trans-acting
determinants of transcription termination by yeast RNA polymerase II. Molec.
Cell. Biol. 26, 2688-2696.
Butcher,
S. and Brow, D.A. (2005). Towards understanding the catalytic core structure of
the spliceosome. Biochem. Soc. Transact.
33, 447-449.
Kwan,
S.S. and D.A. Brow. (2005). The N- and C-terminal RNA recognition motifs of
splicing factor Prp24 have distinct functions in U6 RNA binding. RNA 11,
808-820.
Kaiser,
M.W., J. Chi and D.A. Brow. (2004). Position-dependent function of a B block
promoter element implies a specialized chromatin structure on the S. cerevisiae U6 RNA gene, SNR6. Nucl. Acids Res. 32, 4297-4305.
Professor of Biomolecular Chemistry
Ph.D., University of California at San Diego, 1986
Postdoctoral Research: University of California at San Francisco
Address: 4204B Biochemical Sciences
Telephone: 262-1475
E-mail: dabrow@wisc.edu
Research Interests:
Nuclear steps in eukaryotic gene expression
Research Fields:
Gene Expression
Yeast and Fungi
We investigate the molecular mechanisms of eukaryotic gene expression, with emphasis on transcription and pre-mRNA splicing. Because the fundamental mechanisms of gene expression are conserved among eukaryotes, we have chosen the genetically tractable yeast S. cerevisiae as a model system. Our transcription studies focus on how RNA polymerases (Pol) II and III identify a gene, and on the detailed mechanism of initiation and termination of transcription by Pol II. We use genetic, genomic, and biochemical approaches in these studies. For example, we have measured Pol II occupancy across the entire yeast genome at 100-200 base-pair resolution by chromatin immunoprecipitation, and we have identified mutations in Pol II that cause it to “read-through” transcription terminators. We discovered and are characterizing a Pol II termination pathway that uses the helicase Sen1 and appears to be most important for short transcripts. In our splicing studies we examine the mechanism and function of dynamic RNA-RNA and RNA-protein interactions in assembling the active site of the spliceosome. We defined a complex network of genetic interactions that involve two RNAs (U4 and U6), two helicases, an RNA-binding protein, and the largest and most conserved spliceosomal protein (Prp8). We use detailed mutational studies, in vitro splicing and RNA-binding assays, and structural biology to dissect the mechanism of spliceosome activation. Mutations in the human Prp8 and Sen1 genes cause retinal degeneration and progressive motor neuron dysfunction, respectively. We hope our studies will illuminate the mechanisms of these diseases.
Brow, D.A.
(2011). Sen-sing RNA terminators. Molec. Cell 42, 717-718. Martin-Tumasz,
S., Richie, A.C., Clos, L.J. II, Brow†, D.A., and Butcher†,
S.E. (2011). A novel occluded RNA
recognition motif in Prp24 unwinds the U6 RNA internal stem loop. Nucl.
Acids Res. doi: 10.1093/nar/gkr455.
†corresponding authors Brow,
D.A. Eye on RNA unwinding. (2009). Nature Struct. Molec. Biol. 16, 7-8. Kuehner,
J.N. and Brow, D.A. Regulation of a eukaryotic gene
by GTP-dependent start site selection and transcription attenuation. (2008).
Molec. Cell 31, 201-211. (Cover
article) McManus,
C.J., Schwartz, M.L., Butcher, S.E. and Brow, D.A. (2007). A dynamic bulge in
the U6 RNA internal stem-loop functions in spliceosome assembly and activation.
RNA 13, 2252-2265. Bae, E.,
N.J. Reiter, C.A. Bingman, S.S. Kwan, D. Lee, G.N. Phillips Jr., S.E. Butcher,
and D.A. Brow. (2007). Structure and interactions of the first three RNA
recognition motifs of splicing factor Prp24. J. Mol. Biol., 1447-1458. Steinmetz,
E.J., C.L. Warren, J.N. Kuehner, B. Panbehi, A.Z. Ansari, and D.A. Brow.
(2006). Genome-wide distribution of yeast RNA polymerase II and its control by
Sen1 helicase. Molec. Cell 24,
735-746. (Cover article) Kuehner,
J.N, and D.A. Brow. (2006). Quantitative analysis of in vivo initiator selection by yeast RNA polymerase II supports a
scanning model. J. Biol. Chem. 281,
14119-14128. Steinmetz,
E.J., S.B.H. Ng, J.P. Cloute, and D.A. Brow. (2006). Cis- and trans-acting
determinants of transcription termination by yeast RNA polymerase II. Molec.
Cell. Biol. 26, 2688-2696. Butcher,
S. and Brow, D.A. (2005). Towards understanding the catalytic core structure of
the spliceosome. Biochem. Soc. Transact.
33, 447-449. Kwan,
S.S. and D.A. Brow. (2005). The N- and C-terminal RNA recognition motifs of
splicing factor Prp24 have distinct functions in U6 RNA binding. RNA 11,
808-820. Kaiser,
M.W., J. Chi and D.A. Brow. (2004). Position-dependent function of a B block
promoter element implies a specialized chromatin structure on the S. cerevisiae U6 RNA gene, SNR6. Nucl. Acids Res. 32, 4297-4305.
