Profile

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. . . .

Publications

The following timeline graph is generated from all co-authored publications.

Featured publications are shown below:

  1. ETAA1 acts at stalled replication forks to maintain genome integrity. Bass TE, Luzwick JW, Kavanaugh G, Carroll C, Dungrawala H, Glick GG, Feldkamp MD, Putney R, Chazin WJ, Cortez D (2016) Nat Cell Biol 18(11): 1185-1195
    › Primary publication · 27723720 (PubMed) · PMC5245861 (PubMed Central)
  2. Novel CPVT-Associated Calmodulin Mutation in CALM3 (CALM3-A103V) Activates Arrhythmogenic Ca Waves and Sparks. Gomez-Hurtado N, Boczek NJ, Kryshtal DO, Johnson CN, Sun J, Nitu FR, Cornea RL, Chazin WJ, Calvert ML, Tester DJ, Ackerman MJ, Knollmann BC (2016) Circ Arrhythm Electrophysiol 9(8)
    › Primary publication · 27516456 (PubMed) · PMC4988333 (PubMed Central)
  3. Novel calmodulin mutations associated with congenital long QT syndrome affect calcium current in human cardiomyocytes. Pipilas DC, Johnson CN, Webster G, Schlaepfer J, Fellmann F, Sekarski N, Wren LM, Ogorodnik KV, Chazin DM, Chazin WJ, Crotti L, Bhuiyan ZA, George AL (2016) Heart Rhythm 13(10): 2012-9
    › Primary publication · 27374306 (PubMed) · PMC5035189 (PubMed Central)
  4. Spectrum and Prevalence of CALM1-, CALM2-, and CALM3-Encoded Calmodulin Variants in Long QT Syndrome and Functional Characterization of a Novel Long QT Syndrome-Associated Calmodulin Missense Variant, E141G. Boczek NJ, Gomez-Hurtado N, Ye D, Calvert ML, Tester DJ, Kryshtal DO, Hwang HS, Johnson CN, Chazin WJ, Loporcaro CG, Shah M, Papez AL, Lau YR, Kanter R, Knollmann BC, Ackerman MJ (2016) Circ Cardiovasc Genet 9(2): 136-46
    › Primary publication · 26969752 (PubMed) · PMC4907364 (PubMed Central)
  5. Identification and Optimization of Anthranilic Acid Based Inhibitors of Replication Protein A. Patrone JD, Pelz NF, Bates BS, Souza-Fagundes EM, Vangamudi B, Camper DV, Kuznetsov AG, Browning CF, Feldkamp MD, Frank AO, Gilston BA, Olejniczak ET, Rossanese OW, Waterson AG, Chazin WJ, Fesik SW (2016) ChemMedChem 11(8): 893-9
    › Primary publication · 26748787 (PubMed) · PMC4838552 (PubMed Central)
  6. Promotion of BRCA2-Dependent Homologous Recombination by DSS1 via RPA Targeting and DNA Mimicry. Zhao W, Vaithiyalingam S, San Filippo J, Maranon DG, Jimenez-Sainz J, Fontenay GV, Kwon Y, Leung SG, Lu L, Jensen RB, Chazin WJ, Wiese C, Sung P (2015) Mol Cell 59(2): 176-87
    › Primary publication · 26145171 (PubMed) · PMC4506714 (PubMed Central)
  7. Biochemical and Proteomic Analysis of Ubiquitination of Hsc70 and Hsp70 by the E3 Ligase CHIP. Soss SE, Rose KL, Hill S, Jouan S, Chazin WJ (2015) PLoS One 10(5): e0128240
    › Primary publication · 26010904 (PubMed) · PMC4444009 (PubMed Central)
  8. Functional dynamics in replication protein A DNA binding and protein recruitment domains. Brosey CA, Soss SE, Brooks S, Yan C, Ivanov I, Dorai K, Chazin WJ (2015) Structure 23(6): 1028-38
    › Primary publication · 26004442 (PubMed) · PMC4456234 (PubMed Central)
  9. Simian virus Large T antigen interacts with the N-terminal domain of the 70 kD subunit of Replication Protein A in the same mode as multiple DNA damage response factors. Ning B, Feldkamp MD, Cortez D, Chazin WJ, Friedman KL, Fanning E (2015) PLoS One 10(2): e0116093
    › Primary publication · 25706313 (PubMed) · PMC4337903 (PubMed Central)
  10. Diphenylpyrazoles as replication protein a inhibitors. Waterson AG, Kennedy JP, Patrone JD, Pelz NF, Feldkamp MD, Frank AO, Vangamudi B, Souza-Fagundes EM, Rossanese OW, Chazin WJ, Fesik SW (2015) ACS Med Chem Lett 6(2): 140-5
    › Primary publication · 25699140 (PubMed) · PMC4329576 (PubMed Central)
  11. Dimerization and phosphatase activity of calcyclin-binding protein/Siah-1 interacting protein: the influence of oxidative stress. Topolska-Woś AM, Shell SM, Kilańczyk E, Szczepanowski RH, Chazin WJ, Filipek A (2015) FASEB J 29(5): 1711-24
    › Primary publication · 25609429 (PubMed) · PMC4415008 (PubMed Central)
  12. Human PrimPol is a highly error-prone polymerase regulated by single-stranded DNA binding proteins. Guilliam TA, Jozwiakowski SK, Ehlinger A, Barnes RP, Rudd SG, Bailey LJ, Skehel JM, Eckert KA, Chazin WJ, Doherty AJ (2015) Nucleic Acids Res 43(2): 1056-68
    › Primary publication · 25550423 (PubMed) · PMC4333378 (PubMed Central)
  13. Redefining the DNA-binding domain of human XPA. Sugitani N, Shell SM, Soss SE, Chazin WJ (2014) J Am Chem Soc 136(31): 10830-3
    › Primary publication · 25056193 (PubMed) · PMC4132956 (PubMed Central)
  14. Novel calmodulin mutations associated with congenital arrhythmia susceptibility. Makita N, Yagihara N, Crotti L, Johnson CN, Beckmann BM, Roh MS, Shigemizu D, Lichtner P, Ishikawa T, Aiba T, Homfray T, Behr ER, Klug D, Denjoy I, Mastantuono E, Theisen D, Tsunoda T, Satake W, Toda T, Nakagawa H, Tsuji Y, Tsuchiya T, Yamamoto H, Miyamoto Y, Endo N, Kimura A, Ozaki K, Motomura H, Suda K, Tanaka T, Schwartz PJ, Meitinger T, Kääb S, Guicheney P, Shimizu W, Bhuiyan ZA, Watanabe H, Chazin WJ, George AL (2014) Circ Cardiovasc Genet 7(4): 466-74
    › Primary publication · 24917665 (PubMed) · PMC4140998 (PubMed Central)
  15. Structural analysis of replication protein A recruitment of the DNA damage response protein SMARCAL1. Feldkamp MD, Mason AC, Eichman BF, Chazin WJ (2014) Biochemistry 53(18): 3052-61
    › Primary publication · 24730652 (PubMed) · PMC4020579 (PubMed Central)
  16. Discovery of a potent stapled helix peptide that binds to the 70N domain of replication protein A. Frank AO, Vangamudi B, Feldkamp MD, Souza-Fagundes EM, Luzwick JW, Cortez D, Olejniczak ET, Waterson AG, Rossanese OW, Chazin WJ, Fesik SW (2014) J Med Chem 57(6): 2455-61
    › Primary publication · 24491171 (PubMed) · PMC3969094 (PubMed Central)
  17. Insights into eukaryotic primer synthesis from structures of the p48 subunit of human DNA primase. Vaithiyalingam S, Arnett DR, Aggarwal A, Eichman BF, Fanning E, Chazin WJ (2014) J Mol Biol 426(3): 558-69
    › Primary publication · 24239947 (PubMed) · PMC3946992 (PubMed Central)
  18. Discovery of a potent inhibitor of replication protein a protein-protein interactions using a fragment-linking approach. Frank AO, Feldkamp MD, Kennedy JP, Waterson AG, Pelz NF, Patrone JD, Vangamudi B, Camper DV, Rossanese OW, Chazin WJ, Fesik SW (2013) J Med Chem 56(22): 9242-50
    › Primary publication · 24147804 (PubMed) · PMC3932990 (PubMed Central)
  19. Xeroderma pigmentosum complementation group C protein (XPC) serves as a general sensor of damaged DNA. Shell SM, Hawkins EK, Tsai MS, Hlaing AS, Rizzo CJ, Chazin WJ (2013) DNA Repair (Amst) 12(11): 947-53
    › Primary publication · 24051049 (PubMed) · PMC3825762 (PubMed Central)
  20. Surface reengineering of RPA70N enables cocrystallization with an inhibitor of the replication protein A interaction motif of ATR interacting protein. Feldkamp MD, Frank AO, Kennedy JP, Patrone JD, Vangamudi B, Waterson AG, Fesik SW, Chazin WJ (2013) Biochemistry 52(37): 6515-24
    › Primary publication · 23962067 (PubMed) · PMC3804075 (PubMed Central)
  21. Discovery of Protein-Protein Interaction Inhibitors of Replication Protein A. Patrone JD, Kennedy JP, Frank AO, Feldkamp MD, Vangamudi B, Pelz NF, Rossanese OW, Waterson AG, Chazin WJ, Fesik SW (2013) ACS Med Chem Lett 4(7): 601-605
    › Primary publication · 23914285 (PubMed) · PMC3728914 (PubMed Central)
  22. Stable RAGE-heparan sulfate complexes are essential for signal transduction. Xu D, Young JH, Krahn JM, Song D, Corbett KD, Chazin WJ, Pedersen LC, Esko JD (2013) ACS Chem Biol 8(7): 1611-20
    › Primary publication · 23679870 (PubMed) · PMC3806902 (PubMed Central)
  23. Activation of UbcH5c~Ub is the result of a shift in interdomain motions of the conjugate bound to U-box E3 ligase E4B. Soss SE, Klevit RE, Chazin WJ (2013) Biochemistry 52(17): 2991-9
    › Primary publication · 23550736 (PubMed) · PMC3666176 (PubMed Central)
  24. Calmodulin mutations associated with recurrent cardiac arrest in infants. Crotti L, Johnson CN, Graf E, De Ferrari GM, Cuneo BF, Ovadia M, Papagiannis J, Feldkamp MD, Rathi SG, Kunic JD, Pedrazzini M, Wieland T, Lichtner P, Beckmann BM, Clark T, Shaffer C, Benson DW, Kääb S, Meitinger T, Strom TM, Chazin WJ, Schwartz PJ, George AL (2013) Circulation 127(9): 1009-17
    › Primary publication · 23388215 (PubMed) · PMC3834768 (PubMed Central)
  25. A new structural framework for integrating replication protein A into DNA processing machinery. Brosey CA, Yan C, Tsutakawa SE, Heller WT, Rambo RP, Tainer JA, Ivanov I, Chazin WJ (2013) Nucleic Acids Res 41(4): 2313-27
    › Primary publication · 23303776 (PubMed) · PMC3575853 (PubMed Central)
  26. Lysophosphatidic acid targets vascular and oncogenic pathways via RAGE signaling. Rai V, Touré F, Chitayat S, Pei R, Song F, Li Q, Zhang J, Rosario R, Ramasamy R, Chazin WJ, Schmidt AM (2012) J Exp Med 209(13): 2339-50
    › Primary publication · 23209312 (PubMed) · PMC3526353 (PubMed Central)
  27. Sample preparation methods to analyze DNA-induced structural changes in replication protein A. Brosey CA, Tsutakawa SE, Chazin WJ (2012) Methods Mol Biol : 101-22
    › Primary publication · 22976179 (PubMed) · PMC3713622 (PubMed Central)
  28. Structure of an E3:E2~Ub complex reveals an allosteric mechanism shared among RING/U-box ligases. Pruneda JN, Littlefield PJ, Soss SE, Nordquist KA, Chazin WJ, Brzovic PS, Klevit RE (2012) Mol Cell 47(6): 933-42
    › Primary publication · 22885007 (PubMed) · PMC3462262 (PubMed Central)
  29. Zinc sequestration by the neutrophil protein calprotectin enhances Salmonella growth in the inflamed gut. Liu JZ, Jellbauer S, Poe AJ, Ton V, Pesciaroli M, Kehl-Fie TE, Restrepo NA, Hosking MP, Edwards RA, Battistoni A, Pasquali P, Lane TE, Chazin WJ, Vogl T, Roth J, Skaar EP, Raffatellu M (2012) Cell Host Microbe 11(3): 227-39
    › Primary publication · 22423963 (PubMed) · PMC3308348 (PubMed Central)
  30. Binding to retinoblastoma pocket domain does not alter the inter-domain flexibility of the J domain of SV40 large T antigen. Williams CK, Vaithiyalingam S, Hammel M, Pipas J, Chazin WJ (2012) Arch Biochem Biophys 518(2): 111-8
    › Primary publication · 22227098 (PubMed) · PMC3279518 (PubMed Central)
  31. A high-throughput fluorescence polarization anisotropy assay for the 70N domain of replication protein A. Souza-Fagundes EM, Frank AO, Feldkamp MD, Dorset DC, Chazin WJ, Rossanese OW, Olejniczak ET, Fesik SW (2012) Anal Biochem 421(2): 742-9
    › Primary publication · 22197419 (PubMed) · PMC3274598 (PubMed Central)
  32. Human DNA helicase B (HDHB) binds to replication protein A and facilitates cellular recovery from replication stress. Guler GD, Liu H, Vaithiyalingam S, Arnett DR, Kremmer E, Chazin WJ, Fanning E (2012) J Biol Chem 287(9): 6469-81
    › Primary publication · 22194613 (PubMed) · PMC3307258 (PubMed Central)
  33. BID binds to replication protein A and stimulates ATR function following replicative stress. Liu Y, Vaithiyalingam S, Shi Q, Chazin WJ, Zinkel SS (2011) Mol Cell Biol 31(21): 4298-309
    › Primary publication · 21859891 (PubMed) · PMC3209332 (PubMed Central)
  34. E2 conjugating enzyme selectivity and requirements for function of the E3 ubiquitin ligase CHIP. Soss SE, Yue Y, Dhe-Paganon S, Chazin WJ (2011) J Biol Chem 286(24): 21277-86
    › Primary publication · 21518764 (PubMed) · PMC3122187 (PubMed Central)
  35. Structural basis for ligand recognition and activation of RAGE. Koch M, Chitayat S, Dattilo BM, Schiefner A, Diez J, Chazin WJ, Fritz G (2010) Structure 18(10): 1342-52
    › Primary publication · 20947022 (PubMed) · PMC4250094 (PubMed Central)
  36. Insights into eukaryotic DNA priming from the structure and functional interactions of the 4Fe-4S cluster domain of human DNA primase. Vaithiyalingam S, Warren EM, Eichman BF, Chazin WJ (2010) Proc Natl Acad Sci U S A 107(31): 13684-9
    › Primary publication · 20643958 (PubMed) · PMC2922289 (PubMed Central)
  37. Solution NMR structure of the C-terminal DNA binding domain of Mcm10 reveals a conserved MCM motif. Robertson PD, Chagot B, Chazin WJ, Eichman BF (2010) J Biol Chem 285(30): 22942-9
    › Primary publication · 20489205 (PubMed) · PMC2906287 (PubMed Central)
  38. Structure of a DNA polymerase alpha-primase domain that docks on the SV40 helicase and activates the viral primosome. Huang H, Weiner BE, Zhang H, Fuller BE, Gao Y, Wile BM, Zhao K, Arnett DR, Chazin WJ, Fanning E (2010) J Biol Chem 285(22): 17112-22
    › Primary publication · 20234039 (PubMed) · PMC2878050 (PubMed Central)
  39. Structural dynamics and single-stranded DNA binding activity of the three N-terminal domains of the large subunit of replication protein A from small angle X-ray scattering. Pretto DI, Tsutakawa S, Brosey CA, Castillo A, Chagot ME, Smith JA, Tainer JA, Chazin WJ (2010) Biochemistry 49(13): 2880-9
    › Primary publication · 20184389 (PubMed) · PMC2847624 (PubMed Central)
  40. Direct ubiquitination of beta-catenin by Siah-1 and regulation by the exchange factor TBL1. Dimitrova YN, Li J, Lee YT, Rios-Esteves J, Friedman DB, Choi HJ, Weis WI, Wang CY, Chazin WJ (2010) J Biol Chem 285(18): 13507-16
    › Primary publication · 20181957 (PubMed) · PMC2859511 (PubMed Central)
  41. S100A8/A9 induces autophagy and apoptosis via ROS-mediated cross-talk between mitochondria and lysosomes that involves BNIP3. Ghavami S, Eshragi M, Ande SR, Chazin WJ, Klonisch T, Halayko AJ, McNeill KD, Hashemi M, Kerkhoff C, Los M (2010) Cell Res 20(3): 314-31
    › Primary publication · 19935772 (PubMed) · PMC4161879 (PubMed Central)
  42. S100A8/A9: a Janus-faced molecule in cancer therapy and tumorgenesis. Ghavami S, Chitayat S, Hashemi M, Eshraghi M, Chazin WJ, Halayko AJ, Kerkhoff C (2009) Eur J Pharmacol 625(1-3): 73-83
    › Primary publication · 19835859 (PubMed)
  43. Replication protein A stimulates the Werner syndrome protein branch migration activity. Sowd G, Wang H, Pretto D, Chazin WJ, Opresko PL (2009) J Biol Chem 284(50): 34682-91
    › Primary publication · 19812417 (PubMed) · PMC2787331 (PubMed Central)
  44. Engineering a ubiquitin ligase reveals conformational flexibility required for ubiquitin transfer. Qian SB, Waldron L, Choudhary N, Klevit RE, Chazin WJ, Patterson C (2009) J Biol Chem 284(39): 26797-802
    › Primary publication · 19648119 (PubMed) · PMC2785368 (PubMed Central)
  45. Physical interactions between Mcm10, DNA, and DNA polymerase alpha. Warren EM, Huang H, Fanning E, Chazin WJ, Eichman BF (2009) J Biol Chem 284(36): 24662-72
    › Primary publication · 19608746 (PubMed) · PMC2782055 (PubMed Central)
  46. Checkpoint kinase ATR promotes nucleotide excision repair of UV-induced DNA damage via physical interaction with xeroderma pigmentosum group A. Shell SM, Li Z, Shkriabai N, Kvaratskhelia M, Brosey C, Serrano MA, Chazin WJ, Musich PR, Zou Y (2009) J Biol Chem 284(36): 24213-22
    › Primary publication · 19586908 (PubMed) · PMC2782015 (PubMed Central)
  47. Structural basis for DNA binding by replication initiator Mcm10. Warren EM, Vaithiyalingam S, Haworth J, Greer B, Bielinsky AK, Chazin WJ, Eichman BF (2008) Structure 16(12): 1892-901
    › Primary publication · 19081065 (PubMed) · PMC2636851 (PubMed Central)
  48. Subversion of antimicrobial calprotectin (S100A8/S100A9 complex) in the cytoplasm of TR146 epithelial cells after invasion by Listeria monocytogenes. Zaia AA, Sappington KJ, Nisapakultorn K, Chazin WJ, Dietrich EA, Ross KF, Herzberg MC (2009) Mucosal Immunol 2(1): 43-53
    › Primary publication · 19079333 (PubMed)
  49. The basic cleft of RPA70N binds multiple checkpoint proteins, including RAD9, to regulate ATR signaling. Xu X, Vaithiyalingam S, Glick GG, Mordes DA, Chazin WJ, Cortez D (2008) Mol Cell Biol 28(24): 7345-53
    › Primary publication · 18936170 (PubMed) · PMC2593429 (PubMed Central)
  50. Structure of the S100A6 complex with a fragment from the C-terminal domain of Siah-1 interacting protein: a novel mode for S100 protein target recognition. Lee YT, Dimitrova YN, Schneider G, Ridenour WB, Bhattacharya S, Soss SE, Caprioli RM, Filipek A, Chazin WJ (2008) Biochemistry 47(41): 10921-32
    › Primary publication · 18803400 (PubMed) · PMC2666074 (PubMed Central)
  51. S100A8/A9 at low concentration promotes tumor cell growth via RAGE ligation and MAP kinase-dependent pathway. Ghavami S, Rashedi I, Dattilo BM, Eshraghi M, Chazin WJ, Hashemi M, Wesselborg S, Kerkhoff C, Los M (2008) J Leukoc Biol 83(6): 1484-92
    › Primary publication · 18339893 (PubMed) · PMC2737328 (PubMed Central)
  52. S100A8/9 induces cell death via a novel, RAGE-independent pathway that involves selective release of Smac/DIABLO and Omi/HtrA2. Ghavami S, Kerkhoff C, Chazin WJ, Kadkhoda K, Xiao W, Zuse A, Hashemi M, Eshraghi M, Schulze-Osthoff K, Klonisch T, Los M (2008) Biochim Biophys Acta 1783(2): 297-311
    › Primary publication · 18060880 (PubMed)