Walter J. Chazin, Ph.D.
Chancellor's Prof Biochemistry and Physics
Director, Vanderbilt University Center for Structural Biology
Ingram Professor of Cancer Research
Vanderbilt University Medical Center
5140 BioSci/MRB III
Nashville, TN 37232
Structural Cell Biology- DNA replication, recombination and repair, ubiquitination, calcium signal transduction
1. DNA REPLICATION, RECOMBINATION AND REPAIR.
Our program in structural cell biology includes research to understand the mechanisms of action of multi-protein assemblies that process DNA. Current efforts are focused on establishing and understanding the various functional roles of replication protein A (RPA), a heterotrimeric protein of 70, 32 and 14 kDa subunits. RPA is the major single-strand DNA binding protein in the cell and has been shown to form an integral part of most DNA processing assemblies. Our studies involve proteomic mapping of interactions with other DNA processing proteins and 3D structure determination of protein and protein-DNA complexes. These studies are providing novel insights into the structural basis for mutational defects leading to cancer and the essential foundation for the development of anticancer therapies.
2. PROTEIN UBIQUITINATION
Signalling via protein ubiquitination plays an essential role in an increasing number of cellular processes. A series of ligases catalyze the activation and subsequent covalent attachment of ubiquitin to substrate proteins. The E3 class of ubiquitin ligases typically involves the coordinated activity of a group of proteins that form a multi-protein complex of substrate and E2 ligase recognition elements, scaffolds, and molecular bridges. Our reseach involves the study of a newly discovered ubiquitin ligase activity associated with the RNA spliceosome and of a novel calcium-regulated ubiquitination process that is required for proper embryogenesis and development. Our studies involve proteomic strategies to determine which members of the complex interact with eachother and 3D structure determination of the corresponding protein complexes. These results are revealing information about how protein ubiquitination functions as a signaling mechanism and how defects in E3 ligase proteins lead to malfunction in fundamental cellular processes.
3. CALCIUM SIGNAL TRANSDUCTION
Calcium and calcium-binding proteins (CaBPs) play a central role in intracellular signal transduction pathways and are associated with a wide-range of effects on health and disease. Our research is directed towards understanding the molecular basis for the cellular activities of CaBPs, particularly the transduction of calcium signals to cellular targets. The primary experimental approach involves the determination of 3D structures of specific CaBPs in the presence and absence of calcium and cellular targets, using NMR, X-ray crystallography and computational methods. These are complemented by protein engineering and small molecule design. Ultimately, we seek the ability to control binding properties and interactions with target proteins, thereby enabling the design of specific biological activities and therapeutic strategies relevant to CaBP-mediated disease. . . .
- Chazin, WJ Evolution of the NIGMS Protein Structure Initiative. Structure, 16(1), 12-4, 2008.
- Corbin, BD, Seeley, EH, Raab, A, Feldmann, J, Miller, MR, Torres, VJ, Anderson, KL, Dattilo, BM, Dunman, PM, Gerads, R, Caprioli, RM, Nacken, W, Chazin, WJ, Skaar, EP Metal chelation and inhibition of bacterial growth in tissue abscesses. Science, 319(5865), 962-5, 2008.
- Weiner, BE, Huang, H, Dattilo, BM, Nilges, MJ, Fanning, E, Chazin, WJ An Iron-Sulfur Cluster in the C-terminal Domain of the p58 Subunit of Human DNA Primase. J Biol Chem, 282(46), 33444-51, 2007.
- Ball, HL, Ehrhardt, MR, Mordes, DA, Glick, GG, Chazin, WJ, Cortez, D Function of a Conserved Checkpoint Recruitment Domain in ATRIP Proteins. Mol Cell Biol, 2007.
- Dattilo, BM, Fritz, G, Leclerc, E, Kooi, CW, Heizmann, CW, Chazin, WJ The extracellular region of the receptor for advanced glycation end products is composed of two independent structural units. Biochemistry, 46(23), 6957-70, 2007.
- Chazin, WJ The impact of X-ray crystallography and NMR on intracellular calcium signal transduction by EF-hand proteins: crossing the threshold from structure to biology and medicine. Sci STKE, 2007(388), pe27, 2007.
- Bunick, CG, Miller, MR, Fuller, BE, Fanning, E, Chazin, WJ Biochemical and structural domain analysis of xeroderma pigmentosum complementation group C protein. Biochemistry, 45(50), 14965-79, 2006.
- Shah, VN, Wingo, TL, Weiss, KL, Williams, CK, Balser, JR, Chazin, WJ Calcium-dependent regulation of the voltage-gated sodium channel hH1: Intrinsic and extrinsic sensors use a common molecular switch. Proc Natl Acad Sci U S A, 2006.
- Johnson, E, Chazin, WJ, Rance, M Effects of calcium binding on the side-chain methyl dynamics of calbindin D9k: a 2H NMR relaxation study. J Mol Biol, 357(4), 1237-52, 2006.
- Jiang, X, Klimovich, V, Arunkumar, AI, Hysinger, EB, Wang, Y, Ott, RD, Guler, GD, Weiner, B, Chazin, WJ, Fanning, E Structural mechanism of RPA loading on DNA during activation of a simple pre-replication complex. EMBO J, 25(23), 5516-26, 2006.
- Sheehan, JH, Bunick, CG, Hu, H, Fagan, PA, Meyn, SM, Chazin, WJ Structure of the N-terminal calcium sensor domain of centrin reveals the biochemical basis for domain-specific function. J Biol Chem, 281(5), 2876-81, 2006.
- Vander Kooi, CW, Ohi, MD, Rosenberg, JA, Oldham, ML, Newcomer, ME, Gould, KL, Chazin, WJ The Prp19 U-box crystal structure suggests a common dimeric architecture for a class of oligomeric E3 ubiquitin ligases. Biochemistry, 45(1), 121-30, 2006.
- Sunahori, K, Yamamura, M, Yamana, J, Takasugi, K, Kawashima, M, Yamamoto, H, Chazin, WJ, Nakatani, Y, Yui, S, Makino, H The S100A8/A9 heterodimer amplifies proinflammatory cytokine production by macrophages via activation of nuclear factor kappa B and p38 mitogen-activated protein kinase in rheumatoid arthritis. Arthritis Res Ther, 8(3), R69, 2006.
- Meyn, SM, Seda, C, Campbell, M, Weiss, KL, Hu, H, Pastrana-Rios, B, Chazin, WJ The biochemical effect of Ser167 phosphorylation on Chlamydomonas reinhardtii centrin. Biochem Biophys Res Commun, 342(1), 342-8, 2006.
- Malmendal, A, Vander Kooi, CW, Nielsen, NC, Chazin, WJ Calcium-Modulated S100 Protein-Phospholipid Interactions. An NMR Study of Calbindin D(9k) and DPC. Biochemistry, 44(17), 6502-6512, 2005.
- Arunkumar, AI, Klimovich, V, Jiang, X, Ott, RD, Mizoue, L, Fanning, E, Chazin, WJ Insights into hRPA32 C-terminal domain-mediated assembly of the simian virus 40 replisome. Nat Struct Mol Biol, 12(4), 332-9, 2005.
- Nakatani, Y, Yamazaki, M, Chazin, WJ, Yui, S Regulation of S100A8/A9 (calprotectin) binding to tumor cells by zinc ion and its implication for apoptosis-inducing activity. Mediators Inflamm, 2005(5), 280-92, 2005.
- Bhattacharya, S, Lee, YT, Michowski, W, Jastrzebska, B, Filipek, A, Kuznicki, J, Chazin, WJ The modular structure of SIP facilitates its role in stabilizing multiprotein assemblies. Biochemistry, 44(27), 9462-71, 2005.
- Wingo, Tammy L, Shah, Vikas N, Anderson, Mark E, Lybrand, Terry P, Chazin, Walter J, Balser, Jeffrey R An EF-hand in the sodium channel couples intracellular calcium to cardiac excitability. Nat Struct Mol Biol, 11(3), 219-25, 2004.
- Bunick, Christopher G, Nelson, Melanie R, Mangahas, Sheryll, Hunter, Michael J, Sheehan, Jonathan H, Mizoue, Laura S, Bunick, Gerard J, Chazin, Walter J Designing sequence to control protein function in an EF-hand protein. J Am Chem Soc, 126(19), 5990-8, 2004.
- Christodoulou, John, Malmendal, Anders, Harper, Jeffrey F, Chazin, Walter J Evidence for differing roles for each lobe of the calmodulin-like domain in a calcium-dependent protein kinase. J Biol Chem, 279(28), 29092-100, 2004.
- Lee, Young-Tae, Jacob, Jaison, Michowski, Wojciech, Nowotny, Marcin, Kuznicki, Jacek, Chazin, Walter J Human Sgt1 binds HSP90 through the CHORD-Sgt1 domain and not the tetratricopeptide repeat domain. J Biol Chem, 279(16), 16511-7, 2004.
- Stauffer, Melissa E, Chazin, Walter J Physical interaction between replication protein A and Rad51 promotes exchange on single-stranded DNA. J Biol Chem, 279(24), 25638-45, 2004.
- Stauffer, Melissa E, Chazin, Walter J Structural mechanisms of DNA replication, repair, and recombination. J Biol Chem, 279(30), 30915-8, 2004.
- Bhattacharya, Shibani, Bunick, Christopher G, Chazin, Walter J Target selectivity in EF-hand calcium binding proteins. Biochim Biophys Acta, 1742(1-3), 69-79, 2004.
- Hu, Haitao, Sheehan, Jonathan H, Chazin, Walter J The mode of action of centrin. Binding of Ca2+ and a peptide fragment of Kar1p to the C-terminal domain. J Biol Chem, 279(49), 50895-903, 2004.
- Arunkumar, AI, Stauffer, ME, Bochkareva, E, Bochkarev, A, Chazin, WJ Independent and coordinated functions of replication protein A tandem high affinity single-stranded DNA binding domains. J Biol Chem, 278(42), 41077-82, 2003.
- Ohi, Melanie D, Vander Kooi, Craig W, Rosenberg, Joshua A, Chazin, Walter J, Gould, Kathleen L Structural insights into the U-box, a domain associated with multi-ubiquitination. Nat Struct Biol, 10250-5, 2003.
- Bhattacharya, S, Large, E, Heizmann, CW, Hemmings, B, Chazin, WJ Structure of the Ca2+/S100B/NDR kinase peptide complex: insights into S100 target specificity and activation of the kinase. Biochemistry, 42(49), 14416-26, 2003.
- Hu, H, Chazin, WJ Unique features in the C-terminal domain provide caltractin with target specificity. J Mol Biol, 330(3), 473-84, 2003.
- MÃ¤ler, L, Blankenship, J, Rance, M, Chazin, W J Site-site communication in the EF-hand Ca2+-binding protein calbindin D9k. Nat Struct Biol, 7(3), 245-50, 2000.
- Mer, G, Bochkarev, A, Gupta, R, Bochkareva, E, Frappier, L, Ingles, C J, Edwards, A M, Chazin, W J Structural basis for the recognition of DNA repair proteins UNG2, XPA, and RAD52 by replication factor RPA. Cell, 103(3), 449-56, 2000.
- Smith, JA, Bifulco, G, Case, DA, Boger, DL, Gomez-Paloma, L, Chazin, WJ The structural basis for in situ activation of DNA alkylation by duocarmycin SA. J Mol Biol, 300(5), 1195-204, 2000.
- Botuyan, M V, Koth, C M, Mer, G, Chakrabartty, A, Conaway, J W, Conaway, R C, Edwards, A M, Arrowsmith, C H, Chazin, W J Binding of elongin A or a von Hippel-Lindau peptide stabilizes the structure of yeast elongin C. Proc Natl Acad Sci U S A, 96(16), 9033-8, 1999.
- Miick, S M, Fee, R S, Millar, D P, Chazin, W J Crossover isomer bias is the primary sequence-dependent property of immobilized Holliday junctions. Proc Natl Acad Sci U S A, 94(17), 9080-4, 1997.
- Linse, S, JÃ¶nsson, B, Chazin, W J The effect of protein concentration on ion binding. Proc Natl Acad Sci U S A, 92(11), 4748-52, 1995.
- Potts, B C, Smith, J, Akke, M, Macke, T J, Okazaki, K, Hidaka, H, Case, D A, Chazin, W J The structure of calcyclin reveals a novel homodimeric fold for S100 Ca(2+)-binding proteins. Nat Struct Biol, 2(9), 790-6, 1995.
- Skelton, N J, KÃ¶rdel, J, Akke, M, ForsÃ©n, S, Chazin, W J Signal transduction versus buffering activity in Ca(2+)-binding proteins. Nat Struct Biol, 1(4), 239-45, 1994.