John H. Bushweller
- BA, Dartmouth College
- PhD, University of California, Berkeley, CA
- Postdoc, Eidgenössische Technische Hochschule, Zürich, Switzerland
- Professor, Molecular Physiology and Biological Physics
Structural and Functional Basis for Oncogenesis; Targeted Drug Development; Structural Studies of Membrane Proteins
Structural and Functional Basis for Oncogenesis.
Our lab is fundamentally interested in understanding, from a structural and biophysical perspective, the functioning of proteins involved in regulating transcription, particularly those involved in the dysregulation associated with the development of cancer. Structural and functional characterization of the native forms of these proteins and their relevant complexes via NMR spectroscopy, X-ray crystallography, and a variety of other techniques provides a baseline of understanding. Subsequent characterization of the oncoprotein forms then provides a detailed understanding of the molecular mechanism of oncogenesis associated with altered forms of these proteins. Such knowledge leads to novel avenues for the design of therapeutic agents to treat the cancers associated with these particular oncoproteins. Our current focus is structural studies of a novel transcriptional enhancer referred to as the core-binding factor (CBF). This heterodimeric protein is essential for hematopoietic development. Gene translocations associated with the genes coding for the two subunits of CBF produce novel fusion proteins which have been implicated as playing a role in more than 30% of acute leukemias. We are carrying out structural and functional studies of the oncoprotein forms of the two subunits of CBF that are associated with leukemia. We have now extended these studies to the MLL protein, a key epigenetic regulator that is the target of chromosomal translocations in leukemia which are particularly poor prognosis.
Structure-Based Drug Discovery.
We are using structure-based drug design to develop small molecule inhibitors of the oncoprotein forms of core binding factor as well as of MLL fusion proteins. Our focus is on the development of highly targeted molecules which inactivate the oncoprotein form and have minimal side-effects. Such agents have significant potential for the treatment of the associated leukemias.
Structural Studies of Membrane Proteins.
A third focus for the lab is the application of solution NMR methods to the structure determination of membrane proteins. The vast majority of drug targets are membrane-embedded proteins. This class of proteins has presented significant challenges for structure determination by any method. We determined the structure of the 4 TM enzyme DsbB by solution NMR which established a paradigm for tackling this class of proteins by NMR. We are currently examining additional technical improvements in this area as well as targeting several new systems for structure determination.
Please see our group website for additional information: http://www.people.virginia.edu/~jhb4v/
- Regan M, Horanyi P, Pryor E, Sarver J, Cafiso D, Bushweller J. Structural and dynamic studies of the transcription factor ERG reveal DNA binding is allosterically autoinhibited. Proceedings of the National Academy of Sciences of the United States of America. 2013;110(33): 13374-9. PMID: 23898196 | PMCID: PMC3746864
- Leach B, Kuntimaddi A, Schmidt C, Cierpicki T, Johnson S, Bushweller J. Leukemia fusion target AF9 is an intrinsically disordered transcriptional regulator that recruits multiple partners via coupled folding and binding. Structure (London, England : 1993). 2012;21(1): 176-83. PMID: 23260655 | PMCID: PMC3545106
- Corpora T, Roudaia L, Oo Z, Chen W, Manuylova E, Cai X, Chen M, Cierpicki T, Speck N, Bushweller J. Structure of the AML1-ETO NHR3-PKA(RIIα) complex and its contribution to AML1-ETO activity. Journal of molecular biology. 2010;402(3): 560-77. PMID: 20708017 | PMCID: PMC2945414
- Kamikubo Y, Zhao L, Wunderlich M, Corpora T, Hyde R, Paul T, Kundu M, Garrett L, Compton S, Huang G, Wolff L, Ito Y, Bushweller J, Mulloy J, Liu P. Accelerated leukemogenesis by truncated CBF beta-SMMHC defective in high-affinity binding with RUNX1. Cancer cell. 2010;17(5): 455-68. PMID: 20478528 | PMCID: PMC2874204
- Park S, Osmers U, Raman G, Schwantes R, Diaz M, Bushweller J. The PHD3 domain of MLL acts as a CYP33-regulated switch between MLL-mediated activation and repression . Biochemistry. 2010;49(31): 6576-86. PMID: 20677832 | PMCID: PMC2916634
- Cierpicki T, Bielnicki J, Zheng M, Gruszczyk J, Kasterka M, Petoukhov M, Zhang A, Fernandez E, Svergun D, Derewenda U, Bushweller J, Derewenda Z. The solution structure and dynamics of the DH-PH module of PDZRhoGEF in isolation and in complex with nucleotide-free RhoA. Protein science : a publication of the Protein Society. 2009;18(10): 2067-79. PMID: 19670212 | PMCID: PMC2786971
- Cierpicki T, Risner L, Grembecka J, Lukasik S, Popovic R, Omonkowska M, Shultis D, Zeleznik-Le N, Bushweller J. Structure of the MLL CXXC domain-DNA complex and its functional role in MLL-AF9 leukemia. Nature structural & molecular biology. 2009;17(1): 62-8. PMID: 20010842 | PMCID: PMC2908503
- Park S, Chen W, Cierpicki T, Tonelli M, Cai X, Speck N, Bushweller J. Structure of the AML1-ETO eTAFH domain-HEB peptide complex and its contribution to AML1-ETO activity. Blood. 2009;113(15): 3558-67. PMID: 19204326 | PMCID: PMC2668852
- Park S, Speck N, Bushweller J. The role of CBFbeta in AML1-ETO's activity. Blood. 2009;114(13): 2849-50. PMID: 19779050
- Roudaia L, Cheney M, Manuylova E, Chen W, Morrow M, Park S, Lee C, Kaur P, Williams O, Bushweller J, Speck N. CBFbeta is critical for AML1-ETO and TEL-AML1 activity. Blood. 2009;113(13): 3070-9. PMID: 19179469 | PMCID: PMC2662647
- Zheng M, Cierpicki T, Momotani K, Artamonov M, Derewenda U, Bushweller J, Somlyo A, Derewenda Z. On the mechanism of autoinhibition of the RhoA-specific nucleotide exchange factor PDZRhoGEF. BMC structural biology. 2009;9 36. PMID: 19460155 | PMCID: PMC2695464
- Erfurth F, Popovic R, Grembecka J, Cierpicki T, Theisler C, Xia Z, Stuart T, Diaz M, Bushweller J, Zeleznik-Le N. MLL protects CpG clusters from methylation within the Hoxa9 gene, maintaining transcript expression. Proceedings of the National Academy of Sciences of the United States of America. 2008;105(21): 7517-22. PMID: 18483194 | PMCID: PMC2396713
- Zhou Y, Cierpicki T, Jimenez R, Lukasik S, Ellena J, Cafiso D, Kadokura H, Beckwith J, Bushweller J. NMR solution structure of the integral membrane enzyme DsbB: functional insights into DsbB-catalyzed disulfide bond formation. Molecular cell. 2008;31(6): 896-908. PMID: 18922471 | PMCID: PMC2622435
- Gorczynski M, Grembecka J, Zhou Y, Kong Y, Roudaia L, Douvas M, Newman M, Bielnicka I, Baber G, Corpora T, Shi J, Sridharan M, Lilien R, Donald B, Speck N, Brown M, Bushweller J. Allosteric inhibition of the protein-protein interaction between the leukemia-associated proteins Runx1 and CBFbeta. Chemistry & biology. 2007;14(10): 1186-97. PMID: 17961830
- Liu Y, Chen W, Gaudet J, Cheney M, Roudaia L, Cierpicki T, Klet R, Hartman K, Laue T, Speck N, Bushweller J. Structural basis for recognition of SMRT/N-CoR by the MYND domain and its contribution to AML1/ETO's activity. Cancer cell. 2007;11(6): 483-97. PMID: 17560331 | PMCID: PMC1978186
- Matheny C, Speck M, Cushing P, Zhou Y, Corpora T, Regan M, Newman M, Roudaia L, Speck C, Gu T, Griffey S, Bushweller J, Speck N. Disease mutations in RUNX1 and RUNX2 create nonfunctional, dominant-negative, or hypomorphic alleles. The EMBO journal. 2007;26(4): 1163-75. PMID: 17290219 | PMCID: PMC1852839
- Cierpicki T, Kim M, Cooper D, Derewenda U, Bushweller J, Derewenda Z. The DC-module of doublecortin: dynamics, domain boundaries, and functional implications. Proteins. 2006;64(4): 874-82. PMID: 16835924
- Cierpicki T, Liang B, Tamm L, Bushweller J. Increasing the accuracy of solution NMR structures of membrane proteins by application of residual dipolar couplings. High-resolution structure of outer membrane protein A. Journal of the American Chemical Society. 2006;128(21): 6947-51. PMID: 16719475 | PMCID: PMC2527590
- Grembecka J, Cierpicki T, Devedjiev Y, Derewenda U, Kang B, Bushweller J, Derewenda Z. The binding of the PDZ tandem of syntenin to target proteins. Biochemistry. 2006;45(11): 3674-83. PMID: 16533050
- Li Z, Lukasik S, Liu Y, Grembecka J, Bielnicka I, Bushweller J, Speck N. A mutation in the S-switch region of the Runt domain alters the dynamics of an allosteric network responsible for CBFbeta regulation. Journal of molecular biology. 2006;364(5): 1073-83. PMID: 17059830 | PMCID: PMC1783549
- Liang B, Bushweller J, Tamm L. Site-directed parallel spin-labeling and paramagnetic relaxation enhancement in structure determination of membrane proteins by solution NMR spectroscopy. Journal of the American Chemical Society. 2006;128(13): 4389-97. PMID: 16569016 | PMCID: PMC3199951
- Liu Y, Cheney M, Gaudet J, Chruszcz M, Lukasik S, Sugiyama D, Lary J, Cole J, Dauter Z, Minor W, Speck N, Bushweller J. The tetramer structure of the Nervy homology two domain, NHR2, is critical for AML1/ETO's activity. Cancer cell. 2006;9(4): 249-60. PMID: 16616331