CURRICULUM VITAE JOHN H. LAITY Associate Professor, Director of SBS NMR Facility, School of Biological Sciences University of Missouri-Kansas City 109 Biological Sciences Building Kansas City, MO 64110-2499 Ph: (816) 235-5345/Fax: (816) 235-6584/email: [email protected] EDUCATION The Scripps Research Institute, La Jolla, CA Cornell University, Ithaca, NY Cornell University, Ithaca, NY Columbia University, New York, NY Rutgers University, New Brunswick, NJ

2001 1997 1997 1988 1986

Postdoctoral Fellow, NMR Postdoctoral, Biophysics Ph.D., Chemistry M.S., Chemistry B.S., Chemistry

TEACHING AND RESEARCH Academic Positions: Associate Professor, School of Biological Sciences University of Missouri-Kansas City Assistant Professor, School of Biological Sciences University of Missouri-Kansas City Non-Academic Positions: Director, UMKC, School of Biological Sciences High field NMR Facility

2008- present

2001-2008

2001-Present

Courses Taught: University of Missouri-Kansas City: LS401 Biophysical Principles (course coordinator) Fall 2001-2008 LSMBB565 Protein Structure and Function Winter 2003, 2005, 2007 LSCBB569 Structural Biology, Methods and Strategies Fall 2006 LSMBB596/597 Advanced Experimental Cell Biology 2002-present LSCBB596/597 Advanced Experimental Cell Biology 2002-present LS399/499F Introduction to Research 2003-present Ten UMKC undergraduate students have worked in the Laity research lab while enrolled in LS399/499 since 2003. Five of these students have subsequently been authors on at least one peer-reviewed journal article as a result of their research efforts (undergraduate authors are denoted with * in publication list). Cornell University: Chem203/204 General Chemistry Laboratory

1989-1990

Columbia University: Graduate Course Biophysical Chemistry Physical Chemistry Laboratory

1988 1986-1987

SCHOLASTIC HONORS AND SCHOLARSHIPS UMKC Trustees Faculty Scholar Award National Institutes of Health (NIH) Postdoctoral Fellowship Keystone Millennium Conference Travel Award TSRI Society of Fellows Conference Travel Award NIH Predoctoral Biotechnology Fellowship NIH Predoctoral Biophysics Fellowship Rutgers University Chemistry Department High Honors Rutgers University Chemistry Merck Index Award

2007 1998-2001 2000 2001 1993-1995 1990-1993 1986 1986

PROFESSIONAL SOCIETIES AND ACTIVITIES Professional Organizations: American Association for the Advancement of Science American Chemical Society American Society for Biochemistry and Molecular Biology Protein Society FASEB Society State and National Grant Review Service: NIH Study Section, MSFA study section, at-large panel member, 2007, 2008 University of Missouri Research Board, grant reviewer 2002-2004 Petroleum Research Fund (American Chemical Society), grant reviewer 2005 Editorial Boards and Review: Editor, Open Magnetic Resonance Journal, 2007-present Reviewer, Journal of Molecular Biology, 2001-present Reviewer, Protein Science, 2004-present Reviewer, Proteins: Structure, Function, and Bioinformatics, 2004-Present Reviewer, Biochemistry, 2004-present Reviewer, Journal of Biological Chemistry, 2006-Present Reviewer, Proceedings of the National Academy of Sciences (USA), 2007-Present Reviewer, Structural and Functional Genomics, 2006-Present Reviewer, Human Molecular Genetics, 2007-Present Reviewer, Journal of Biomedical Discover and Collaboration, 2007- Present Reviewer, Journal of Molecular Graphics and Modeling, 2004 Reviewer, Chemical Research in Toxicology, 2004

RESEARCH AREAS OF INTEREST NMR is a well-suited technique to investigate structural and dynamic changes of proteins in solution. As the home of the School of Biological Sciences (SBS) NMR Facility, we are using NMR and other biophysical and molecular biology approaches to explore several aspects of zinc finger transcription factor biology. A major focus of the lab is to characterize the molecular mechanisms that control divalent zinc [Zn(II)] homeostasis in yeast and mammalian cells. All known organisms require Zn(II), and yet this essential nutrient is toxic if accumulated to excess. Zn(II) homeostasis in mammalian and yeast cells is regulated at least in part at the level of coordination chemistry by two zinc finger proteins, Zap1 (yeast) and MTF-1 (mammalian). MTF-1 and Zap1 are transcription factors that control the expression of proteins to transport Zn(II) in or out of the cellular compartments [Zn(II) transport proteins] or to sequester Zn(II) through high-affinity metal-ligand coordination (metallothioneins). Studies on MTF-1 involve a close collaboration with Dr. Glen Andrews at the neighboring University of Kansas Medical Center, Department of Molecular Biology and Biochemistry. A new area of focus in the lab centers on understanding the molecular mechanisms whereby a special group of zinc finger-containing Zic family transcription factors regulate key processes that are essential for human brain development. These studies are ongoing in close collaboration with Dr. Gerald Wyckoff in the SBS. In two additional collaborative projects, we are exploring the molecular mechanisms of targeted elements of bacterial pathogenesis. We are currently collaborating with Dr. Lawrence Dreyfus, Dean of the UMKC, SBS to explore the structure/function relationships that underlie the cytotoxic effects of the cytolethal distending toxin. In a second collaboration with Dr. Brian Geisbrecht (SBS), we are investigating structural and dynamic aspects of antigen/receptor interactions. RESEARCH SUPPORT (Past 4 years) Ongoing R01 GM073771 Period: 04/01/07-03/31/12 NIH/NIGMS $$301,514 current year ($1,517,534 total award) “Mechanisms of Eukaryotic Zinc Homeostasis” Role: PI (Co-I, Andrews, G.K.) R03 AI69001 Period: 06/01/06-05/31/08 NIH/NIAID $50,000 direct current year ($100,000 total direct) “Structural Studies of the MPT64 Family of Mycobacterial Virulence Factors” Role: Co-I (PI, Geisbrecht, B.V.) R15 HD055668 Period: 04/15/08-03/31/11 NIH/NICHD $150,000 direct current year ($221,050 total award) “Role of ZIC and GLI Proteins in Human Brain Disorders” Role: Co-I (PI, Wyckoff, G.J.) Completed 3055 Period: 01/01/06-12/31/06 University of Missouri Research Board (UMRB) $30,000 direct to JHL

“Characterization of the MTF-1 Metalloregulatory Domain” Role: PI S10 RR022341 Period: one-time 04/010/06 NIH/NIGMS $500,000 direct (shared instrumentation grant). “800 MHz Spectrometer for Biomolecular NMR in Missouri” Role: Co-I (PI, Van Doren, S) R01 ES05704 NIH/NIEHS $82,755 direct to JHL “Environmental Toxicology Using Transgenic Models” Role: Co-I (PI, Andrews, G.K.)

Period: 08/08/02-07/31/05

University of Missouri SEARCH Period: 09/01/05-12/31/05 UMKC SEARCH fund $800 direct “NMR Structural and Dynamic Characterization of the Wild-Type and Disulfide Mutant Forms of the MPT64 Protein from Mycobacterium Tuberculosis” Role: Faculty mentor University of Missouri SEARCH Period: 09/01/05-12/31/05 UMKC SEARCH fund $1500 direct “Structural and Functional Evolution of Interacting Zinc Finger Proteins” Role: Faculty mentor 1197 UMRB “Molecular Mechanisms of Zinc Regulation.” Role: PI

Period: 02/01/03-1/31/04 $41,058 direct to JHL

ADMINISTRATIVE EXPERIENCE Academic: Member, Interdiscipinary Ph.D. Executive Committee (Campus-wide), Biological and Chemical Science Subcommittee, University of Missouri-Kansas City, 2008-2010. Member, Cell Biology and Biophysics Division Search Committee, University of MissouriKansas City, 2002, 2004. Member, Structural Biology/Biotechnology Committee, 2002-present. Member, Administrative Issues Committee, University of Missouri-Kansas City, 2003. Member, Information Technologies Committee, School of Biological Sciences, 2003-present. INTERNATIONAL MEETINGS Poster: “Comparison of Backbone Structures and Thermodynamic Stabilities of Wild-Type and [C65A, C72A] Bovine Pancreatic Ribonuclease A: Heteronuclear NMR Analysis of a Disulfide Folding Intermediate”, International Conference on Protein Folding and Design, 1996 NATIONAL MEETINGS

Poster Presentations and abstracts: “NMR studies of structure-regulatory relationships in DNA binding and nuclear export functions involving interacting Zic1 C2H2 zinc fingers”, Keystone Symposia-Frontiers in Structural Biology, Steamboat Springs, CO, 2008.

“NMR solution structure and DNA binding heterogeneity of the 6-zinc-finger DNA-binding and metalloregulatory domain of MTF-1”, Keystone Symposia-Frontiers in Structural Biology, Steamboat Springs, CO, 2008. “Structural and Functional Evolution of Interacting C2H2 Zinc Finger Proteins”, Experimental Biology 2006: Advancing the Biomedical Frontier, San Francisco, CA, 2006. “Structure and Solution Dynamics of CdtB, the Biologically Active Subunit of Cytolethal Distending Toxin”, Experimental Biology 2006: Advancing the Biomedical Frontier San Francisco, CA, 2006. “NMR Structural and Dynamic Studies of the Six Cys2His2 Zinc Fingers of MTF-1 in the Free and DNA-Bound State”, FASEB Summer Research Conference- Trace Element Metabolism: Integrating Basic and Applied Approach. Snowmass, CO, 2004. “Metal-Sensing Mechanisms of Two Zap1 Zinc Fingers”, FASEB Summer Research Conference- Trace Element Metabolism: Integrating Basic and Applied Approach, Snowmass, CO, 2004. “New Paradigms for Zinc Finger Structure and Function”, Keystone Symposia-Frontiers in Structural Biology, Snowbird, UT, 2004. “ Molecular Mechanisms of Zinc Homeostasis Probed by NMR”, Keystone Symposia-Frontiers of NMR in Molecular Biology, Taos, NM, 2003. “ Molecular Basis for Differing Biological Roles of Alternative Splice Forms of the Wilms’ Tumor Suppressor Protein”, Keystone Symposia-Frontiers of NMR in Molecular Biology, Bozeman, MT, 2001. “ Iterative Splicing in Wilms’ Tumor Suppressor Protein is a New Paradigm of Cellular Regulation and Control of Oncogenesis”, Keystone Symposia-Millennium Conference, Keystone, CO, 2000. "NMR Studies of DNA Binding in Alternative Splice Forms of Wilms' Tumor Suppressor Protein", Keystone Symposia-Frontiers of NMR in Molecular Biology, Big Sky, MT, 1999. “NMR Studies of Alternative Splicing in Wilms Tumor Suppressor Protein”, 39th Experimental Nuclear Magnetic Resonance Conference, Asilomar, CA, 1998.

“Automated NMR Sequential Backbone Assignments and Structural Characterization of the Major Rate-Determining Disulfide Folding Intermediate Analog [C40A, C95A] Bovine Pancreatic Ribonuclease A”, Keystone Symposia-Frontiers of NMR in Molecular Biology, Taos, NM, 1997. “Heteronuclear NMR Studies of [C65A, C72A] Ribonuclease A. A Model of a Major Oxidative Folding Intermediate”, Protein Science, 4, Suppl. 2, p. 242-S, 1995. “Conformational Studies of Three-Disulfide Mutants of Bovine Pancreatic Ribonuclease A”, Protein Science, 2, Suppl. 1, p. 63, 1993. “Expression of Wild-Type and Mutant Bovine Pancreatic Ribonuclease A”, A.C.S. Meeting Abstracts, 203, p. Biot-28, 1992. Other Meeting Participation: Judge for undergraduate student poster session, American Society for Biochemistry and Molecular Biology 100th Annual Meeting, San Francisco, CA, 2006. INVITED LECTURES 2007 University of Missouri-Kansas City, School of Biological Sciences, Kansas City, MO, “Mechanisms of Eukaryotic Zinc Homeostasis”. 2006 Great Plains Regional Annual Symposium on Protein NMR, Lawrence, KS, “Why is NMR so well suited for identifying and characterizing functionally important interaction sites in Zinc Finger proteins?” 2006 Experimental Biology Symposium 2006, San Francisco, CA (four invited presentations): 1. Advancing the Biomedical Frontier San Francisco, CA, “Structural and Functional Evolution of Interacting C2H2 Zinc Finger Proteins” 2, “Towards a better diagnostic for active tuberculosis: NMR solution structure and mutational engineering of protein Rv1980c from M. tuberculosis” 3. “Molecular Mechanisms of MTF-1 Metalloregulation” 4, “Solution structure of a Zap1 zinc responsive domain provides insights into metalloregulatory transcriptional repression in S. cerevisiae” 2005 4th Annual Great Plains Infectious Disease Meeting, Lawrence, KS, “A Tale of Two Pathogens: NMR Structural and Dynamic Studies of the M. tuberculosis Rv1980c and E.Coli CtdB proteins” 2005 University of Kansas, Department of Molecular Biosciences, Lawrence KS, “How much Zinc is enough? Structural, dynamic, and metal binding studies of two eukaryotic metalloregulatory transcription factors” 2005 University of Kansas Medical Center, Department of Biochemistry and Molecular Biology, Kansas City, KS, “Are We Full of zinc? Binding and structural studies of metalloregulatory domains from MTF-1 and Zap1” 2005 University of Louisville, Department of Chemistry, Louisville, KY,“ Probing the Metal Responsive Transcriptional Mechanisms Underlying Zinc Homeostasis at the Molecular Level”.

2004 2003 2003

2003 2003 2002 2001 2001

2001

2001

2001

2001

2000

2000

COBRE Symposium on Protein Structure and Function, Lawrence, KS, “NMR Studies of Metal Sensing Zinc Fingers”. University of Missouri-Columbia, Department of Chemistry, Kansas City, MO, “New Paradigms of Zinc Finger Structure and Function” University of Missouri-Kansas City, School of Dentistry, Kansas City, MO “Probing the Molecular Details of Specificity and Function in Cys2-His2 Zinc Finger Transcription Factors” University of Kansas Medical Center, Department of Biochemistry and Molecular Biology, Kansas City, KS, “title” Kansas State University, Department of Chemistry, Manhattan, Kansas, “New Paradigms of Zinc Finger Structure and Function” University of Missouri-Kansas City, Association of Pharmacy Students, Kansas City, MO “Using high field NMR to explore molecular structure and interactions” University of Missouri-Kansas City, Department of Chemistry, Kansas City, MO, “Using High Field NMR to Explore the Molecular Mechanisms of Transcription” University of Chicago, Department of Chemistry, Chicago, IL, “The Wilms’ Tumor Suppressor Protein: New Insights into DNA Binding and Functional Diversity in Cys2His2 Zinc Fingers” Purdue University, Department of Medicinal Chemistry and Molecular Pharmacology, West Lafayette, IN, “The Wilms’ Tumor Suppressor Protein: New Insights into DNA Binding and Functional Diversity in Cys2His2 Zinc Fingers” University of Nebraska-Lincoln, Department of Chemistry, Lincoln, NE, “The Wilms’ Tumor Suppressor Protein: New Insights into DNA Binding and Functional Diversity in Cys2His2 Zinc Fingers” Washington State University, Department of Chemistry, Pullman, WA, “The Wilms’ Tumor Suppressor Protein: New Insights into DNA Binding and Functional Diversity in Cys2His2 Zinc Fingers” Yale University, Department of Molecular Biophysics and Biochemistry, New Haven, CT, “The Wilms’ Tumor Suppressor Protein: New Insights into DNA Binding and Functional Diversity in Cys2His2 Zinc Fingers” Duke University, Department of Biochemistry, Durham, NC, “The Wilms’ Tumor Supressor Protein: New Insights into DNA Binding and Functional Diversity in Cys2His2 Zinc Fingers ” University of Missouri-Kansas City, School of Biological Sciences, Kansas City, MO, “The Wilms’ Tumor Suppressor Protein: New Insights into DNA Binding and Functional Diversity in Cys2His2 Zinc Fingers ”

THESIS DIRECTION Ph.D. Theses Directed: Student Paul Vordtriede Subhashree Rangarajan Timothy Priddy Zhonghua Wang Elbert Schmitt

Graduation date May 2006 January 2007 January 2007 May 2008 anticipated 2008

(Mentor or Committee member) Committee member Committee member Committee member Mentor Committee member

David Razafsky anticipated 2008 Committee member Ryan Huyck anticipated 2011 Mentor Karen Khan anticipated 2012 Committee member M.S. Theses Directed: Student Graduation date (Mentor or Committee member) Krishna Venkataraman January 2003 Mentor Thesis title: “Study of the Zap1 Zinc Finger Protein Using Nuclear Magnetic Resonance: Software Tools and Methods” Priya Parasuram May 2005 Mentor Thesis Title:“Homologous Modeling and Structural Calculation of the MTF-1 Zinc Finger domain Using NMR and Computation Techniques”

PEER-REVIEWED PUBLICATIONS

1. Li Y, Kimura T, Huyck, RW, Laity JH, Andrews GK. “Zinc-induced formation of a coactivator complex containing the zinc-sensing transcription factor MTF-1, CBP/p300 and Sp1”, Mol Cell Biol, Epub May 5, 2008; 28 (13),p.4275-4284. 2. Stoll, R Lee, BM, Debler, EW, Laity, JH Wilson, IA, Dyson, HJ, and Wright, PE. “Structure of the Wilms Tumor Suppressor Protein Zinc Finger Domain Bound to DNA”, Journal of Molecular Biology, 2007, Oct 5; 372(5), p1227-1245. 3. Wang Z, Potter BM*, Gray AM*, Sacksteder KA, Geisbrecht BV, Laity JH. “The Solution Structure of Antigen MPT64 from Mycobacterium tuberculosis Defines a New Family of Beta-Grasp Proteins,” Journal of Molecular Biology, 2007 Feb 16;366(2), p.375-381. 4. Li Y, Kimura T, Laity JH, Andrews GK.” The zinc-sensing mechanism of mouse MTF-1 involves linker peptides between the zinc fingers,”Mol Cell Biol. 2006 Aug;26(15):5580-7. 5. Hontz JS, Villar-Lecumberri MT, Potter BM*, Yoder MD, Dreyfus LA, Laity JH. “Differences in crystal and solution structures of the cytolethal distending toxin B subunit: Relevance to nuclear translocation and functional activation,”J Biol Chem., 2006 Sep 1; 281(35):25365-72. 6. Wang Z, Feng LS, Matskevich V, Venkataraman K, Parasuram P, Laity JH., “Solution structure of a Zap1 zinc-responsive domain provides insights into metalloregulatory transcriptional repression in Saccharomyces cerevisiae,” J Mol Biol., 2006 Apr 7;357(4):1167-83. *Reviewed by Faculty of 1000 7. Villar-Lecumberri MT, Potter BM*, Wang Z, Dreyfus LA, Laity JH., “NMR Assignment of the E. coli-II Cytolethal Distending Toxin CdtB Subunit.” J Biomol NMR. 2006 Dec;36 Suppl 5:14.

8. Danahy JM*, Potter BM*, Geisbrecht BV, Laity JH., “NMR assignment of protein Rv1980c from Mycobacterium tuberculosis,”J Biomol NMR. 2005 Sep;33(1):73. 9. Potter, B.M.*, Feng, L.S., Matskevich, V.A., Wilson, J.A.*, Andrews, G.K., and Laity, J.H. “The Six Zinc Fingers of Metal Responsive Element Binding Transcription Factor-1 Form Stable Structures with Relatively Small Differences in Zinc Affinities,” Journal of Biological Chemistry, 2005 Aug 5;280(31):28529-40. 10. Potter, B.M.*, Knudsen, N.A.*, Feng, L.S., Matskevich, V.A., Wilson, J.A.*, Andrews, G.K., and Laity, J.H. “NMR assignment of the six zinc fingers of MTF-1 in the free and DNA bound states,” Journal of Biomolecular NMR, 2005 May;32(1):94. 11. Navon, A., Ittah, V., Laity, J.H., Scheraga, H.A., Haas, E., and Gussakovsky, E.E. “Global and Local Steps in the Thermal Denaturation of Bovine Pancreatic Ribonuclease A”, Biochemistry, 40 (1), p. 93-104, 2001. 12. Laity, J.H., Dyson, H.J., and Wright, P.E. “Molecular Basis for Modulation of Biological Function by Alternate Splicing of the Wilms’ Tumor Suppressor Protein”, Proceedings of the National Academy of Science, U.S.A, 97 (22), p. 11932-35, 2000. 13. Laity, J.H., Chung, J., Dyson, H.J., and Wright, P.E.“Alternative Splicing of Wilms’ Tumor Suppressor Protein Modulates DNA Binding Activity Through Isoform-Specific DNAInduced Conformational Changes”, Biochemistry 39 (18), p. 5341-48, 2000. 14. Laity, J.H., Dyson, H.J., Wright, P.E.“DNA-Induced Alpha-Helix Capping in Conserved Linker Sequences is a Determinant of Binding Affinity in Cys2-His2 Zinc Fingers”, Journal of Molecular Biology, 295 (4), p. 719-27, 2000. 15. Laity, J.H., Montelione, G.T., and Scheraga H.A. “Comparison of Local and Global Stability of an Analog of a Disulfide –Folding Intermediate with those of the Wild-Type Protein in Bovine Pancreatic Ribonuclease A: Identification of Specific Regions of Stable Structure Along the Oxidative Folding Pathway”, Biochemistry 38 (50), p.16432-42, 1999. 16. Pearson, M.A., Karplus, P.A., Dodge, R.W., Laity, J.H. and Scheraga, H.A.“Crystal Structures of Two Mutants that have Implications for the Folding of Bovine Pancreatic Ribonuclease A”, Protein Science, 7 (5), p. 1255-8, 1998. 17. Laity, J.H., Lester, C.C., Shimotakahara, S., Zimmermann, D.E., Montelione, G.T., and Scheraga, H.A. “Structural Characterization of an Analog of the Major Rate-Determining Disulfide Folding Intermediate of Bovine Pancreatic Ribonuclease A”, Biochemistry, 36 (42), p. 12683-99, 1997. 18. Lester, C.C., Xu, X., Laity, J.H., Shimotakahara, S., and Scheraga, H.A “Regeneration Studies of a Two-Disulfide Mutant of RNase A”, Biochemistry, 36 (42), p. 13068-76, 1997.

19. Shimotakahara, S., Rios, C.B., Laity, J.H., Zimmerman, D.E., Scheraga, H.A. and Montelione, G.T. “NMR Structural Analysis of an Analog of an Intermediate Formed in the Rate-Determining Step of One Pathway in the Oxidative Folding of Bovine Pancreatic Ribonuclease A: Automated Analysis of 1H 13C, and 15N Resonance Assignments for WildType and [C65S, C72S] Mutant Forms”, Biochemistry, 36 (23), p. 6915-29, 1997. 20. Dodge, R.W., Laity, J.H., Rothwarf, D.M., Shimotakahara, S., and Scheraga, H.A. “Folding Pathway of Guanidine-Denatured Disulfide-Intact Wild-Type and Mutant Bovine Pancreatic Ribonuclease A”, Journal of Protein Chemistry, 13 (4), p. 409-21, 1994. 21. Laity, J.H., Shimotakahara, S., and Scheraga, H.A.“Expression of Wild-Type and Mutant Bovine Pancreatic Ribonuclease A in Escherichia coli”, Proceedings of The National Academy of Science U.S.A., 90 (2), p. 615-9, 1993. 22. Weiner, M.P., Thannhauser,T.W., Laity, J.H., Benning, M.E., Lee, D.P., and Scheraga,H.A. “Plasmid Purification Using Reverse-Phase High Performance Liquid Chromatography Resin PRP-∞”, Nucleic Acids Research, 16 (16), p. 8185, 1988.

INVITED REVIEWS AND BOOK CHAPTERS 1.

Laity, J.H. and Andrews, G.K. “Understanding the mechanisms of zinc sensing by metalresponsive-element binding transcription factor-1 (MTF-1), Arch. Biochem. & Biophys. 2007, 463, 201-210

2.

Laity, J.H., “Cys2-His2 Zinc Finger Proteins”, in Handbook of Metalloproteins, ed. A. Messerschmidt, 3rd ed., 2004, John Wiley & Sons Press, West Sussex, United Kingdom p. 307-323.

3.

Laity, J.H., Lee, B.M., Wright, P.E. “Zinc finger proteins: new insights into structural and functional diversity,” Curr Opin Struct Biol. 2001 Feb;11(1):39-46.