Curriculum Vitae

John P. Marino Leader, Macromolecular Structure & Function Group National Institute of Standards and Technology Biochemical Science Division Gaithersburg, MD 20899 and Adjunct Associate Professor Institute for Bioscience and Biotechnology Research University of Maryland 9600 Gudelsky Drive, Rockville, MD 20850 Phone: (240)-314-6160 Fax: (240)-314-6255 email: [email protected]; [email protected] http://marinolab.umbi.umd.edu/

RESEARCH INTERESTS My research interests are focused in two principle areas: RNA interactions involved in gene regulation and retroviral replication and G-Protein Coupled Receptor signaling. Our RNA work has focused on studies of RNA loop-loop kissing complexes derived from bacterial plasmid regulatory systems and the HIV-1 retroviral dimerization linkage site, the interaction of these RNA complexes with small auxiliary proteins and the interaction of small molecule ligands with potential RNA drug targets. We have also used fluorinated base analogs to characterize retroviral polypurine RNA:DNA hybrids. Our work related to G-Protein Coupled Receptor (GPCR) signaling has aimed to probe the structural basis for guanine nucleotide exchange in G Protein Coupled Receptor signaling. To advance our studies, we have developed new NMR and fluorescence methods to facilitated characterization of the structure, dynamics and binding kinetics of these macromolecular interactions. Overall, the research in my group is driven by the goal of applying precise and general measurement methods to define the structure, thermodynamics and kinetics of RNA structural folds, RNA-ligand complexes and protein interactions at the molecular level. In addition to gaining fundamental insights into macromolecular function, our research aims to impact the fields of pharmaceutical design and development. EDUCATION AND TRAINING 1989 - 1995

Ph. D., Biophysical Chemistry Department of Chemistry Yale University, New Haven, CT Ph.D. Advisors: Donald M. Crothers & James H. Prestegard

1989 - 1991

M. Phil., Chemistry

Marino, John P.

Department of Chemistry Yale University, New Haven, CT 1985 - 1989

A. B., magna cum laude, Chemistry Princeton University, Princeton, NJ Senior Thesis Advisor: G. Charles Dismukes

PROFESSIONAL RESEARCH EXPERIENCE 07/10 - present

Adjunct Associate Professor (tenure-equivalent appointment) University of Maryland, College Park Institute for Bioscience and Biotechnology Research Rockville, MD.

10/08 - present

Leader, Macromolecular Structure & Function Group University of Maryland Biotechnology Institute Center for Advanced Research in Biotechnology Rockville, MD.

09/09 - present

Graduate Faculty Status Department of Chemistry and Biochemistry University of Maryland College Park, MD.

10/08 – 07/10

Associate Director University of Maryland Biotechnology Institute Center for Advanced Research in Biotechnology Rockville, MD.

10/06 - present

Research Chemist V (Career Appointment) National Institute of Standards and Technology Chemical Sciences and Technology Laboratory Biotechnology Division Gaithersburg, MD.

07/04 – 07/10

Adjunct Associate Professor (tenure-equivalent appointment) University of Maryland Biotechnology Institute Center for Advanced Research in Biotechnology Rockville, MD.

08/01 – 9/06

Research Chemist IV (Career Appointment) National Institute of Standards and Technology Chemical Sciences and Technology Laboratory Biotechnology Division Gaithersburg, MD.

09/98 - present

Graduate Faculty Status

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Program in Molecular and Cell Biology University of Maryland College Park, MD. 08/97 – 06/04

Adjunct Assistant Professor University of Maryland Biotechnology Institute Center for Advanced Research in Biotechnology Rockville, MD.

03/01 - 06/01

Visiting Professor (Spring trimester) Department of Chemistry and Chemical Engineering California Institute of Technology Pasadena, CA.

08/97 - 08/01

Research Chemist IV (Term Appointment) National Institute of Standards and Technology Chemical Sciences and Technology Laboratory Biotechnology Division Gaithersburg, MD.

01/95 - 06/97

Alexander von Humboldt Post-Doctoral Fellow Institut für Organische Chemie Johann Wolfgang Goethe-Universität Frankfurt, Germany Postdoctoral Advisor: Christian Griesinger

06/87 - 08/87

Undergraduate Summer Research (URP) Program Cold Spring Harbor Laboratory Cold Spring Harbor, New York Research Advisor: James R. Feramisco

RESEARCH GRANTS, AWARDS AND FELLOWSHIPS NIST Intramural Awards 2002 - 2006

Co-P.I., NIST Intramural Competence Award entitled “Single Molecule Manipulations and Measurements”

2000 - 2002

Co-P.I., Advanced Technologies Program Intramural Award entitled “Single Molecule Probes”

1997 - 2000

Co-P.I., Advanced Technologies Program Intramural Award entitled “Detection, Quantification and Structure of Novel Nucleic Acid-Protein Complexes using Fluorescence and NMR Spectroscopy”

Extramural Grants

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2006 - 2009

P.I. (subcontractor), NIH/EY R01 grant entitled “Structural Analysis of the Rhodopsin-Transducin Complex”

2005 - 2010

P.I., NIH/NIGMS R01 grant entitled “Structural Basis of RNA Stem-Loop ‘Kissing’ Interactions”

2004 - 2005

P.I. (subcontractor), NIH/EY R01 grant entitled “Structural Studies of Gprotein Coupled Receptors”.

1999 - 2004

P.I., NIH/NIGMS R01 grant entitled “Structural Basis of RNA Stem-Loop ‘Kissing’ Interactions”

2002

P.I., NIH/NCRR S10 Shared Instrumentation Grant entitled “Acquisition of a 600 MHz NMR Cryoprobe”

2001

Participant, Keck Foundation Award entitled “Laboratory of Structural Biology at CARB”

2001 - 2005

Collaborator, NIH/EY R01 grant entitled “Structural Studies of G-Protein Coupled Receptors”

Other Fellowships and Awards 2007

Department of Commerce Silver Medal Award

1995 - 1997

Alexander von Humboldt Post-Doctoral Fellowship

1992 - 1995

National Institutes of Health Research Associate

1991 - 1992

Advanced Physical Chemistry Teaching Fellow

1989 - 1991

National Institutes of Health Biophysical Training Grant Award

1989

Merck Award for Undergraduate Research

1985 - 1989

National Merit Scholar

PROFESSIONAL ACTIVITIES 2009

Member, NIH Center for Scientific Review Study Section. NIH/NCRR study section to review NMR infrastructure grants.

2008

Member: DOE review panel of the Environmental and Molecular Sciences Laboratory, PNNL

2007

Temporary Member, NIH Center for Scientific Review MSF-C Study Section

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Marino, John P.

2005

Member: NIH/NCRR Center for Scientific Review P41 reverse site visit review panel that evaluated the NMRFAM Facility in Madison, WI.

2004

Member, NIH Center for Scientific Review Study Section. NIH/NCRR study section to review NMR infrastructure grants.

2002

Member, NIH Center for Scientific Review Study Section. Participated in a site visit review of the RCMI program project grant at Howard University

2001

Temporary Member, NIH Center for Scientific Review Physical Biochemistry Study Section

2000, 2001

Member, NIH Center for Scientific Review study section. Involved in two sitevisit reviews of the National Stable Isotope Resource at Los Alamos, NM.

2000

Chair, Search Committee for recruiting a CARB NMR Facility Manager

1998 - present

Supervisor, CARB/NIST NMR Facility

1998 - 2002

Coordinator, CARB/NIST NMR User Group Seminar series

1997 - 1998

Organizer, CARB Extramural Seminar series

1999 - 2004

Member, MOCB graduate student thesis committee

1997 - present

Ad-hoc Reviewer: Nucleic Acids Research, Journal of the American Chemical Society, Journal of Molecular Biology, Journal of Magnetic Resonance, Journal of Biomolecular NMR, Journal of Biological Chemistry, Biochemistry

1997- present

Member, RNA Society

1997- present

Member, American Chemical Society

1990 - present

Member, AAAS

TEACHING ACTIVITIES 1998, 2002

Lecturer, Seminar series on Modern NMR Methods in Macromolecular Structure Determination, CARB

2001

Lecturer, Methods for the Determination of the Structure of Molecules, Department of Chemistry and Chemical Engineering, California Institute of Technology

1999 - 2001

Lecturer, Protein Structure & Function, Molecular and Cell Biology course (MOCB 640) of the University of Maryland at College Park RESEARCH MENTORING

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Undergraduate & High School Students 06/10 - 08/10 Theresa Brush (University of Maryland, Shady Grove) 06/09 - 05/10

Utkala Segu (Northwest High School, Germantown, MD)

06/06 - 08/06

Kendall Bourke (Boston College, MA)

06/03 - 08/03

Katherine Briggs (Wheeling Jesuit College, WV)

06/02 - 08/02

Katherine Briggs (Wheeling Jesuit College, WV); Julia Tzeng (Winston Churchill High School, MD)

06/01 - 08/01

Dennis Dowhy (Loyola College of Maryland, MD)

06/99 – 8/99

Joseph Tucker (Swarthmore College, PA)

Graduate Students 09/08 -

Katharine Briggs (University of Maryland, College Park, Molecular and Cell Biology Program)

01/05 – 11/09

Hui-Wen Lee (University of Maryland, College Park, Department of Chemistry and Biochemistry)

10/99 - 12/01

Manuela Rist (NIST Visiting Research Scientist from the Technical University of Munich, Awarded a Ph.D. from the TU-Munich in March, 2003)

Post-doctoral Research Associates 07/08 Andrea Szakal (National Research Council Postdoctoral Research Associate) 01/08 – 05/09

Justyna Mysliwy (Postdoctoral Research Associate); Current: Almac CO., UK

06/07 -

Ravi Aduri (Postdoctoral Research Associate)

01/06 – 12/08

Robert G. Brinson (National Research Council Postdoctoral Research Associate); Current: Research Chemist, NIST-Hollings Marine Lab

09/05 - 08/07

Evi Stuble (National Research Council Postdoctoral Research Associate): Current: Staff Fellow, FDA/CBER

09/02 - 09/04

Peter Yim (NIST Postdoctoral Research Associate appointment with the NIST Physics Laboratory)

09/02 - 09/04

Thomas Bradrick (NIH Postdoctoral Research Associate; Current: Examiner, US Patent & Trademark Office)

05/00 - 07/04

Eric S. DeJong (National Research Council Postdoctoral Research Associate [2000-2002]; Current: Examiner, US Patent & Trademark Office)

primary

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04/01-06/03

Mihaela Rita Mihailescu (Current: Associate Professor., Duquesne University)

09/99 - 12/01

Burkhard Luy (Feodor-Lynen Postdoctoral Fellow, Current: Professor, Universität Karlsruhe)

10/98 - 07/99

Hiroko Shimada (Current: Research Associate, Yale University, CT)

02/98 - 08/99

Karen Lacourciere (Current: Program Officer, NIAID/NIH)

PUBLICATIONS (1994-present): Books and Book Chapters 1. C. Griesinger, M. Hennig, J.P. Marino, B. Reif, C. Richter, H. Schwalbe (1999) Methods for the Determination of Torsion Angle Restraints in Biomacromolecules, Biomolecular NMR, Vol. 16, Eds. L. Berliner and R Krishna, Plenum Press, 259-367. Research Articles 1. Kevin B. Turner, Robert G. Brinson, Hye Young Yi-Brunozzi, John P. Marino, Daniele Fabris and Stuart F.J. Le Grice (2009) SHAMS - Combining Chemical Modification of RNA and Mass Spectrometry to Examine Polypurine Tract-Containing RNA/DNA Hybrids, Nucleic Acids Research, 15(8):1605-13. 2. Brinson RG, Turner KB, Yi-Brunozzi HY, Le Grice SF, Fabris D, Marino JP. (2009) Probing anomalous structural features in polypurine tract-containing RNA-DNA hybrids with neomycin B., Biochemistry, 48(29):6988-97. 3. Lee HW, Briggs KT, Marino JP. (2009) Dissecting structural transitions in the HIV-1 dimerization initiation site RNA using 2-aminopurine fluorescence, Methods, 49(2):118-27. 4. Najmoutin G. Abdulaev, Xiang Mao, Eva Ramon, Tony Ngo, Justyna Mysliwy, John P. Marino, and Kevin D. Ridge (2009) Designing Point Mutants to Detect Structural Coupling in a Heterotrimeric G protein α-subunit by NMR Spectroscopy, Photochemistry & Photobiology, 85(2):431-6. 5. Hye Young Yi-Brunozzi, Robert G. Brinson, Danielle M. Brabazon, Daniela Lener, Stuart F. J. Le Grice, and John P. Marino (2008) High Resolution NMR Analysis of the Conformations of Native and Base Analog Substituted Retroviral and LTR-retrotransposon Polypurine Tract Primers, Chem Biol. 2008 Mar;15(3):254-62. [Accompanying preview/highlight: Megan E. Fitzgerald and Alexander C. Drohat Structural Studies of RNA/DNA Polypurine Tracts Chem Biol. 2008 Mar;15(3), 203-204.] 6. Kevin B. Turner, Robert G. Brinson, Hye Young Yi-Brunozzi, Jennifer T. Mille, Jason W. Rausch, Stuart F.J. Le Grice, John P. Marino, and Daniele Fabris (2008) Structural probing of the HIV-1 polypurine tract RNA:DNA hybrid using classic nucleic acid ligands, Nucl. Acids. Res., 2008 May;36(8):2799-810. [Epub 2008 Apr 9.]

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7. Struble, E. B.; Ladner J. E., Brabazon D. M. and Marino J. P. New crystal structures of ColE1 Rom and variants resulting from mutation of a surface exposed residue: Implications for RNArecognition. Proteins: Structure, Function and Bioinformatics. 2008 Feb 7; [Epub ahead of print]. 8. Zhao, C. Marino JP Synthesis of HIV-1 Ψ-site RNA Sequences with site specific incorporation of the fluorescent base analog 2-aminopurine. Tetrahedron. 2007 April; 63: 3575-3584. 9. N. G. Abdulaev, T. Ngo, E. Ramon, D. M. Brabazon, J. P. Marino and K. D. Ridge (2006) The receptor bound ‘empty pocket’ state of the heterotrimeric G-protein α-subunit is conformationally dynamic. Biochemistry. 45(43):12986-97. 10. Ridge KD, J. P. Marino, T. Ngo, E. Ramon, D. M. Brabazon, and N. G. Abdulaev (2006) NMR Analysis of Rhodopsin-Transducin Interactions. J. Vis. Res., 46(27):4482-92. [Epub 2006 Sep 18]. 11. Ridge KD, Abdulaev NG, Zhang C, Ngo T, Brabazon DM, Marino JP. (2006) Conformational changes associated with receptor stimulated guanine nucleotide exchange in a heterotrimeric Gprotein alpha-subunit: NMR analysis of GTPγS-bound states. J. Biol. Chem. 281(11): 7635-48. 2006 Jan 9; [Epub ahead of print] 12. Dash C, Marino JP, Le Grice SF. (2006) Examining Ty3 polypurine tract structure and function by nucleoside analog interference J. Biol. Chem., 281(5):2773-83. Epub 2005 Nov 22. 13. Yi-Brunozzi HY, Brabazon DM, Lener D, Le Grice SF, Marino JP. (2005) A ribose sugar conformational switch in the LTR-retrotransposon Ty3 polypurine tract-containing RNA/DNA hybrid. J. Am. Chem. Soc. 127(47):16344-5. 14. Abdulaev NG, Ngo T, Zhang C, Dinh A, Brabazon DM, Ridge KD, Marino JP. (2005) Heterotrimeric G-protein a-subunit adopts a "preactivated" conformation when associated with βγ-subunits. J. Biol. Chem., 280(45):38071-80. Epub 2005 Aug 29. 15. N. G. Abdulaev, C. Zhang, A. Dinh, T. Ngo, P. N. Bryan, D. M. Brabazon, J. P. Marino, and K. D. Ridge (2005) Bacterial expression and one-step purification of an isotope-labeled heterotrimeric G-protein α-subunit, J. Biomol. NMR, 32(1), 31-40. 16. P.B. Yim, X. Zhang, E. S. DeJong, J. M. Carroll, J.P. Marino and L.S. Goldner (2005) Single RNA Kissing Complexes Studied by Fluorescence Resonance Energy Transfer, Proceedings of the International Society for Optical Engineering, 6(14), 450-459. 17. T. D. Bradrick and J.P. Marino (2004) Ligand-induced changes in 2-aminopurine fluorescence as a probe for small molecule binding to HIV-1 TAR RNA, RNA, 10, 1459-1468. 18. J. Furrer, F. J. Kramer, J. P. Marino, S.J. Glaser and B. Luy (2004) Homonuclear Hartmann-Hann transfer with reduced relaxation losses by use of MOCCA-XY16 multiple pulse sequence, J. Magn. Reson., 166, 39-46.

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19. M.R. Mihailescu and J. P. Marino (2004) A proton-coupled dynamic conformational switch in the HIV-1 dimerization initiation site kissing complex, Proc. Natl. Acad. Sci., USA., 101(5), 11891194. 20. D. M. Brabazon, N. G. Abdulaev, J. P. Marino, and K. D. Ridge (2003) Evidence for Structural Changes in Carboxyl-terminal Peptides of Transducin α-subunit upon binding a Soluble Mimic of Light-activated Rhodopsin, Biochemistry 42(2), 302-311. 21. B. Luy and J. P. Marino (2003) JE-TROSY: combined J- and TROSY-spectroscopy for the Measurement of One-bond Couplings in Macromolecules, J. Magn. Reson. 163, 92-98. 22. E. S. DeJong, C. Chang, M. K. Gilson and J. P. Marino (2003) Proflavine Acts as a Rev Inhibitor by Targeting the High-Affinity Rev Binding Site of the Rev-Responsive Element of HIV-1, Biochemistry 42(26), 8035-8046. 23. B. Luy, U. Richter, E. S. DeJong, O. W. Sørensen and J. P. Marino (2002) Observation of H-bond Mediated 3hJH2H3 Couplings across Watson-Crick AU Base Pairs in RNA, J. Biomol. NMR 24, 133-142. 24. M. J. Rist and J. P. Marino (2002) Mechanism of Nucleocapsid Protein Catalyzed Structural Isomerization of the Dimerization Initiation Site of HIV-1, Biochemistry 41(50), 14762-14770. 25. Luy, B. & Marino, J.P. (2001) 1H-31P CPMG-Correlated Experiments for the Assignment of Nucleic Acids. J. Am. Chem. Soc. 123(45), 11306-11307. 26. Luy, B. & Marino, J.P. (2001) S3E-E.COSY Methods for the Measurement of JHF and JHH Coupling Constants. J. Magn. Reson. 152, 179-184. 27. Luy, B. & Marino, J.P. (2001) Measurement and Application of 1H-19F Dipolar Couplings in the Structure Determination of 2'-Fluorolabeled RNA. J. Biomol. NMR 20, 39-47. 28. Goldner, L. S., K. D. Weston, W. F. Heinz, J. Hwang, E. S. DeJong and J. P. Marino (2001) Single Molecule Probes, Proceedings of the International Society for Optical Engineering, 4450, 70-80. 29. Rist, M.J. & Marino, J.P. (2001) Quantitative Analysis of an Antisense RNA Loop-Loop Complex using 2-Aminopurine Fluorescence. Nucl. Acids. Res. 29(11), 2401-2408. 30. Lacourciere, K.A., Stivers, J. T., and Marino, J. P. (2000) Mechanism of Neomycin and Rev Peptide Binding to the Rev Responsive Element of HIV-1 as Determined by Fluorescence and NMR Spectroscopy. Biochemistry 39(19), 5630-5641. 31. Luy, B. & Marino, J.P. (2000) Direct evidence for Watson-Crick Base-pairs in a Dynamic Region of RNA Structure. J. Am. Chem. Soc. 122(33), 8095-8096. 32. Richter, C., B. Reif, K. Wörner, S. Quant, J.P. Marino, J.W. Engels, C. Griesinger, H. Schwalbe (1998) Determination of 3J(C4',P) coupling constants in Oligonucleotides by a Quantitative J Method. J. Biomol. NMR 12, 223-230.

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33. Marino, J.P., J. Diener, P. B. Moore and C. Griesinger (1997) Two to Threefold Enhancement of 1 H-13C Methine and Methylene Correlations in Uniformly 13C Labeled RNA via Heteronuclear Multiple Quantum Coherence Transfer. J. Am. Chem. Soc. 119, 7361-7366. 34. Marino, J.P., H. Schwalbe, S.J. Glaser and C. Griesinger (1996) Determination of γ and Stereospecific Assignment of H5' Protons by Measurement of 2J and 3J Coupling Constants in Uniformly 13C Labeled RNA. J. Am. Chem. Soc. 118, 4388-4395. 35. Glaser, S.J., H. Schwalbe, J.P. Marino and C. Griesinger (1996) Directed TOCSY, A Method for Selection of Directed Correlations by Optimal Combinations of Isotropic and Longitudinal Mixing. J. Magn. Reson., Series B 112, 160-180. 36. Zimmer, D.P., D.M. Crothers, J.P. Marino and C. Griesinger (1996) NMR Methods for Determination of Homo- and Heteronuclear Coupling Constants in Uniformly 13C,15N Labeled DNA. Magn. Reson. Chem. 34, 177-185. 37. Marino, J.P., H. Schwalbe, C. Anklin, D.M. Crothers, C. Griesinger (1995) Application of 1 13 31 H, C, P-C,C-TOCSY Experiments to the Sequential Backbone Assignment of RNA Oligonucleotides. J. Biomol. NMR 5, 87-92. 38. Marino, J.P., R.S. Gregorian Jr., G. Csankovzki and D.M. Crothers (1995) Bent Helix Formation between RNA Hairpins with Complementary Loops. Science 268, 1448-1454. 39. Schwalbe, H., J.P. Marino, S.J. Glaser, C. Griesinger (1995) Measurement of H,H-Coupling Constants Associated with ν1, ν2, and ν3 in Uniformly 13C Labeled RNA by HCC-TOCSY-CCH E.COSY. J. Am. Chem. Soc. 117, 7251-7252. 40. Marino, J.P., J.H. Prestegard, D.M. Crothers (1994) Correlation of Adenine H2/H8 Resonances in Uniformly 13C Labeled RNAs by 2D HCCH-TOCSY: A New Tool for 1H Assignment. J. Am. Chem. Soc. 116, 2205-2206. 41. Schwalbe, H., J.P. Marino, G.C. King, R. Wechselberger, W. Bermel, C. Griesinger (1994) Determination of a Complete Set of Coupling Constants in 13C-Labeled Oligonucleotides. J. Biomol. NMR 4, 631-644. 42. Marino, J.P., H. Schwalbe, C. Anklin, W. Bermel, D.M. Crothers, C. Griesinger (1994) A ThreeDimensional Triple Resonance 1H,13C,31P Experiment: Sequential Through-Bond Correlation of Ribose Protons and Intervening Phosphorus Along the RNA Oligonucleotide Backbone. J. Am. Chem. Soc. 116, 6472-6473. 43. Ghose, R., J.P. Marino, K.B. Wiberg, J.H. Prestegard (1994) Dependence of 13C Chemical Shift on Glycosidic Torsional Angles in Ribonucleic Acids. J. Am. Chem. Soc. 116, 8827-8828. Review Articles 1. Daniele Fabris, John P. Marino and Stuart F. J. Le Grice (2009) Review: Revisiting Plus-Strand DNA Synthesis in Retroviruses and Long Terminal Repeat Retrotransposons: Dynamics of Enzyme: Substrate Interactions Viruses, 1(3), 657-67

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2. E.S. DeJong, B. Luy & J.P. Marino (2002) Applications of NMR and Fluorescence in RNA Targeted Drug Design, Curr. Top. Med. Chem. 2, 289-302. 3. M.J. Rist & J.P. Marino (2002) Fluorescent Nucleotide Base Analogs as Probes of Nucleic Acid Structure, Dynamics and Interactions, Curr. Org. Chem. 6, 775-793. 4. J.P. Marino, H. Schwalbe and C. Griesinger (1999) J-Coupling Restraints in RNA Structure Determination. Acc. Chem. Res. 32(7) 614-623. Monographs 1. J.P. Marino (1995) Multi-Dimensional Heteronuclear NMR Structural Studies of RNA Oligonucleotides in Solution: (i) New Methods for Through-Bond Assignment and Characterization of Backbone Resonances (ii) Structural Characterization of Complexes formed between RNA Hairpins with Complementary Loops. Dissertation, Yale University. Patents: 1. John P. Marino and Gary G. Giulian (2008) Harvesting of Processed Carbon Nanotubes, Patent Application submitted September 15, 2008. 2. Marino, J.P., Stivers, J.T., and Lacourciere, K.A. (2001) Rapid Fluorescence Detection of Ligand Binding to Nucleic Acid Drug Targets labeled with Highly Fluorescent Nucleotide Base Analogs, Claims of the non-provisional patent accepted 06/03.

INVITED TALKS AND PROFESSIONAL PAPER PRESENTATIONS (1996-present) 2009 1. Poster Presentation, R. Aduri, K.T. Briggs and J.P. Marino, Elucidation of Key Functional Residues Involved in NCp7 Mediated HIV-1 DIS Maturation, The 7th International Retroviral NC Symposium, University of Minnesota, Minneapolis, Minn., September, 2009. 2. Invited Platform Speaker: G.G. Giulian and J.P. Marino, New ‘Green’, Scalable Approaches for Purification and Size Separation of Carbon Nanotubes, - Environmental Implications and Applications of Nanotechnology, UMass, Amherst, MA, June, 2009. 3. Invited Platform Speaker, G.G. Giulian, K.T. Briggs, and J.P. Marino, “Identification and characterization of lithium/magnesium/phosphate complexes relevant to lithium’s bioactivity” 50th ENC, Asilomar CA, March, 2009. 4. Poster Presentation: A. Berton, K.B. Turner, R.G. Brinson, J.P. Marino, S.F.J. LeGrice and D. Fabris ‘Probing RNA structure by selective 2’ hydroxyl acylation analyzed by mass spectrometry (SHAMS), 57th ASMS Conference on Mass Spectrometry, Philadelphia, PA, June, 2009 2008 1. Invited Seminar: J. P. Marino, Carbon Nanotubes: ‘Green Processing’ and New Applications, UMCP, Gaithersburg, MD. October, 2008.

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2. Seminar: J. P. Marino, Carbon Nanotubes: ‘Green Processing’ and New Applications, Nanotoxicity Working Group, NIST, Gaithersburg, MD. July, 2008. 3. Invited Seminar: J. P. Marino, Carbon Nanotubes: ‘Green Processing’ and New Applications, UMBI Faculty Retreat, Solomon Island, MD. June, 2008. 4. Oral Presentation: J.P. Marino, Grit Shear Length Sorting of Nanotubes, NIST Nanotechnology Fair, NIST, Gaithersburg, April 8, 2008 5. Poster Presentation: Gary G. Giulian and John P. Marino, Grit Shear Length Sorting of Nanotubes, NIST Nanotechnology Fair, NIST, Gaithersburg, April, 2008 6. Guest Lecturer: J.P. Marino, Visual Transduction, Shady Grove Bioengineering Program, Rockville, MD, April, 2008. 7. Invited Seminar: J. P. Marino, Probing the Molecular Mechanism of Heterotrimeric G protein Signaling using High-resolution NMR, UMBC, Department of Chemistry, Baltimore, MD, April, 2008. 8. Poster presentation: Robert G. Brinson, Hye Young Yi-Brunozzi, Danielle M. Brabazon, Stuart F. J. Le Grice, and John P. Marino, Application of 2,4-Difluoro-5-methylbenzene as a Tool for NMR Assignment and Analysis of Oligonucleotides with High Sequence Redundancy” 49th ENC, Asilomar CA, March, 2008. 9. Poster presentation: J. P. Marino, N. G. Abdulaev, E. Ramon, X. Mao, T. Ngo, K. D. Ridge, “Probing the Molecular Mechanism of Heterotrimeric G protein Signaling using High-resolution NMR” 49th ENC, Asilomar CA, March, 2008. 10. Poster presentation: G. G. Giulian, K. T. Briggs, and J. P. Marino, “NMR Evidence for Selective Binding of Lithium to Magnesium bound Nucleotide Di- and Tri-phosphates:ADP, GDP, ATP and GTP” 49th ENC, Asilomar CA, March, 2008. 11. Invited Seminar: J. P. Marino “Evidence for selective binding of lithium to magnesium chelation sites in RNA” NIH RNA Club, Bethesda, MD, January, 2008. 2007 1. Invited seminar: J.P. Marino “Probing Heterotrimeric G-protein Signaling using High-resolution NMR” NIST Physics Laboratory, January, 2007. 2. Poster presentation: H.Y. Yi-Brunozzi, R.G. Brinson D. Brabazon, D. Lener, Stuart F.J. Le Grice & J.P. Marino. Application of 2-4-Difluorotoluene as a Tool for NMR Assignment and Analysis of RNA:DNA PPT Hybrids, Keystone Symposium, Frontiers of NMR in Molecular Biology X, Snowbird, Utah, January, 2007. 3. Poster presentation: J. P. Marino, N. G. Abdulaev, E. Ramon, T. Ngo, D. M. Brabazon, and K.D. Ridge. Probing the Mechanism of Heterotrimeric G-protein Signaling using High-resolution NMR Keystone Symposium, Frontiers of NMR in Molecular Biology X, Snowbird, Utah, January, 2007.

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4. Session Chair: Keystone Symposium, Frontiers of NMR in Molecular Biology X, Snowbird, Utah, January, 2007. 5. Poster presentation: E. Struble, J. Ladner, D. Brabazon, J.P. Marino. New crystal structures of wild type ColE1 Rom and variants resulting from mutation of a surface exposed residue: Implications for RNA-recognition, Annual Biophysical Society Meeting, Baltimore, MD March 2007. 6. Seminar: J. P. Marino “Can nucleotide analogs, aminoglycosides and NMR help us understand how PPT’s function?” Tri-Group Meeting (NIST/UMBC/NIH), CARB, Rockville, MD, February, 2007. 7. Invited Seminar: J. P. Marino “RNA and protein structural requirements in HIV-1 DIS Maturation” ACS National Meeting, Chicago, IL, March, 2007. 8. Poster presentation: N. G. Abdulaev, E. Ramon, T. Ngo, K.D. Ridge, and J. P. Marino. Probing the Molecular Mechanism of Heterotrimeric G-protein Signaling using High-resolution NMR ASBMB Annual Meeting, Washington, DC. April 2007. 9. Invited Seminar: J. P. Marino “RNA and protein structural requirements in HIV-1 DIS Maturation” Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, June, 2007. 10. Invited Seminar: J. P. Marino “RNA and protein structural requirements in HIV-1 DIS Maturation” ACS National Meeting, Boston, MA, August, 2007. 2006 1. Seminar, J. P. Marino, “NCp7 sequence requirements for chaperone activity in HIV-1 DIS maturation” RNA Interest Group, Bethesda, MD, Dec., 2006. 2. Seminar, Gary Giulian, John P. Marino, Neşe Sari, Bani Cipriano, Srini Raghavan, Jeeseong Hwang, and Kim Briggman ‘1H, 13C and 23Na NMR analysis of sodium dodecyl sulfate (SDS) “trapped” in mechanically shortened double walled carbon nanotubes (DWNTs) 3. Seminar, Hye Young Yi-Brunozzi, Robert G. Brinson, Daniele Brabazon, Daniela Lener, Stuart F.J. Le Grice & John. P. Marino., “Communication of Structural Signals in PPT-Containing RNA:DNA Hybrids” RNase H 2006, St. Michaels, MD, September, 2006. 4. Seminar, J. P. Marino “Probing structural changes in the HIV-1 dimerization initiation site kissing dimer during retroviral RNA maturation” 20th Meeting of Groups Studying the Structures of AIDS-Related Systems and Their Application to Targeted Drug Design, Bethesda, MD, June, 2006. 5. Seminar, K D. Ridge, N. G. Abdulaev, Tony Ngo, E Ramon, D M. Brabazon, and J P. Marino “Conformational changes in the R*-bound transducin α-subunit accompanying guanine nucleotide release and uptake” PreARVO Meeting 2006, Ft. Lauderdale, FL, April, 2006

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2005 1. Seminar, J. P. Marino “Probing the Mechanism of Heterotrimeric G-protein signaling using highresolution NMR” Vanderbilt University, Nashville, Tennessee, December, 2005. 2. Seminar, J. P. Marino “Probing the Mechanism of Heterotrimeric G-protein signaling using highresolution NMR” UMBI Research Day, University of Maryland, Shady Grove, November, 2005. 3. Seminar, J. P. Marino “Probing the Mechanism of Heterotrimeric G-protein signaling using highresolution NMR” Health Science Center at the Houston Medical School, Houston, Texas, November, 2005. 4. Seminar, J. P. Marino “Probing the Mechanism of Heterotrimeric G-protein signaling using highresolution NMR” Hollings Marine Laboratory, Charleston, South Carolina, October, 2005. 5. Seminar, J. P. Marino, C. Zhao, M. R. Mihailescu & M. J. Rist, “The role of RNA structure in NCp7 chaperoned HIV-1 maturation” 5th International Retroviral NC Symposium, Montreal, Canada September, 2005. 6. Seminar, H-Y. Yi-Brunozzi, D. M. Brabazon, D. Lener, S. F. J. Le Grice and J. P. Marino “A ribose conformational switch in the LTR-retrotransposon Ty3 polypurine tract containing RNA/DNA hybrid” 19th Meeting of Groups Studying the Structures of AIDS-Related Systems and Their Application to Targeted Drug Design, Bethesda, MD, June, 2005. 7. Seminar, D.M. Brabazon, J.E. Ladner & J. P. Marino, “Modulating RNA-binding properties of a four-helix bundle protein by altering protein conformation,” RNA Interest Group, Bethesda, MD, June, 2005. 8. Seminar, J. P. Marino, D.M. Brabazon, N. G. Abdulaev & K.D. Ridge, “High-resolution NMR studies of GPCR/G-protein Signaling” 5th Annual Biotechnolgy Division Retreat, Shepherdstown, WV, May, 2005. 9. Poster presentation/Selected short talk, J. P. Marino, D.M. Brabazon, N. G. Abdulaev & K.D. Ridge, “High-resolution NMR studies of GPCR/G-protein Signaling” 46th ENC, Providence, RI , April, 2005. 10. Poster presentation, D.M. Brabazon, J.E. Ladner & J. P. Marino, “Modulating RNA-binding properties of a four-helix bundle protein by altering protein conformation” 46th ENC, Providence, RI , April, 2005. 11. Poster presentation, “High Resolution NMR Studies of GPCR/G-protein Signaling” Keystone Symposium, Banff, Canada, January, 2005. 2004 1. Seminar, “RNA and other Large Biomolecules and Interactions” Eastern Analytical Symposium, Piscataway, NJ, November, 2004.

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2. Seminar, “High Resolution NMR Studies of GPCR/G-protein Signaling” NIST/UMCP Biotechnology Symposium, College Park, MD, October, 2004. 3. Seminar, “Identifying Small Molecules that Target Retroviral RNA” Department of Molecular Biology, Cleveland Clinic, Cleveland, OH, October, 2004. 4. Seminar, “The Role of RNA Structure in Maturation of the HIV-1 Dimerization Initiation Site Kissing Dimer” 18th Annual Meeting of the Groups Studying the Structure of AIDS-Related Systems and their Application to Targeted Drug Design, Bethesda, MD, June, 2004. 5. Seminar, “A proton-coupled dynamic conformational switch in the HIV-1 dimerization initiation site kissing complex” NIH RNA Club, Bethesda, MD, May, 2004. 6. Poster presentation, M.R. Mihailescu & J.P. Marino, “A proton-coupled dynamic conformational switch in the HIV-1 dimerization initiation site kissing complex” Keystone Symposium, Snowbird, Utah, April, 2004. 7. Seminar, “Dissecting the mechanism of NCp7 catalyzed structural isomerization of the dimerization initiation site” NIH/NHLB, Bethesda, MD, February, 2004. 8. Poster presentation, M.R. Mihailescu & J.P. Marino “The nucleocapsid catalyzed maturation of the Dimerization Initiation Site of HIV-1 is Modulated by pH-dependent RNA conformational Dynamics” 48th Annual Biophysical Society Meeting, Baltimore, MD February, 2004. 9. Poster presentation, T.D. Bradrick & J.P. Marino “Small molecule binding to HIV-1 TAR RNA as monitored by ligand-induced changes in 2-aminopurine fluorescence” 48th Annual Biophysical Society Meeting, Baltimore, MD February, 2004. 10. Poster presentation, P. Yim, L.S. Goldner, A.M. Bardo, E.S. DeJong & J.P. Marino “Characterization of fluorescence resonance energy transfer between pairs of fluorophores using an RNA ruler” 48th Annual Biophysical Society Meeting, Baltimore, MD February, 2004. 11. Seminar, D.M. Brabazon & N. Abdulaev, J.P. Marino & K.D. Ridge, “Mapping Structural changes in the G protein transducin upon binding a soluble mimic of light-activated rhodopsin” 48th Annual Biophysical Society Meeting, Baltimore, MD February, 2004. 2003 1. Poster presentation, M.R. Mihailescu E.S. DeJong & J.P. Marino, “Mechanism of nucleocapsid protein catalyzed structural isomerization of the Dimerization Initiation Site of HIV-1” Fourth HIV DRP Symposium on Antiviral Drug Resistance, Chantilly, VA, December, 2003. 2. Seminar, “Targeting RNA: Fluorescence and NMR techniques to identify new therapeutic leads” Strategic Research Institute, Nucleic Acid World Summit, Boston, MA September, 2003. 3. Seminar, “HIV-1 Nucleocapsid catalyzed maturation of the Dimerization Initiation Site is modulated by pH-dependent RNA dynamics” AIDS Structural Biology Meeting, NIH, Bethesda, MD, June, 2003.

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4. Poster Presentation, “HIV-1 Nucleocapsid catalyzed maturation of the Dimerizatin Initiation Site is modulated by pH-dependent RNA dynamics” Annual NIST Biotechnology Retreat Shepherdstown, WV, May, 2003. 5. Poster Presentation, “Relaxation-Enhanced TOCSY Multiple Pulse Sequence”, 44th Experimental NMR Conference, Savanna, Georgia, April, 2003. 6. Seminar, “Methods for Discovery and Characterization of lead compounds that target RNA” CUNY-Brooklyn College, Brooklyn, NY, April, 2003. 7. Seminar, “Dissecting the Mechanism of NCp7 catalyzed structural isomerization of the dimerization initiation site” EMBL, Heidelberg, Germany, March, 2003. 8. Seminar, “Dissecting the Mechanism of NCp7 catalyzed structural isomerization of the dimerization initiation site” Technical University, Munich, Germany, March, 2003. 9. Seminar, “Dissecting the Mechanism of NCp7 catalyzed structural isomerization of the dimerization initiation site” Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany, March, 2003. 10. Seminar, “Screening small molecules that target RNA using fluorescence and NMR spectroscopy” Frontiers of NMR in Molecular Biology VIII, Taos, New Mexico, February, 2003. 11. Seminar, “RNA: Tool and target in drug discovery” NRC CSTL Review Panel Technical Seminar, February, 2003. 2002 1. Seminar, “Dissecting the Mechanism of NCp7 catalyzed structural isomerization of the dimerization initiation site” NIH RNA Interest Group, Bethesda, MD, December, 2002. 2. Seminar, “Dissecting the Mechanism of NCp7 catalyzed structural isomerization of the dimerization initiation site” 4th Annual MCB Faculty and Student Retreat of the University of Maryland, Baltimore, MD, December, 2002. 3. Poster Presentation, “Mechanism of nucleocapsid protein catalyzed structural isomerization of the Dimerization Initiation Site of HIV-1” Third HIV DRP Symposium on Antiviral Drug Resistance Chantilly, VA, December, 2002 4. Seminar, “Targeting RNA: Development of Fluorescence and NMR Techniques to Identify Novel Therapeutics” RNA in Drug Development: RNA as a Tool and Target, San Diego, CA, November, 2002 5. Seminar, “RNA Loop-Loop Complexes, Structural Variations on an Antisense Theme” International Conference on Magnetic Resonance in Biological Systems, Toronto, CA, August, 2002.

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6. Seminar, “Drug Discovery and Development” Henry Stewart Conference, Washington, DC, July, 2002. 7. Seminar, “HIV-1 DIS: The Kiss of Death” Department of Chemistry, Yale University, New Haven, CT, June, 2002. 8. Seminar, “Targeting Retroviral RNA: Development of Fluorescence and NMR Techniques to Identify Novel Therapeutics” NIH AIDS Structural Biology Meeting, Bethesda, MD, June, 2002. 9. Seminar, “Targeting RNA: Development of Fluorescence and NMR Techniques to Identify Novel Therapeutics” Annual NIST Biotechnology Retreat, Sheperdstown, WV, May, 2002. 10. Seminar, “Mechanism of NCp7 Chaperoned Structural Isomerization of the Dimerization initiation site of HIV-1” Department of Pharmacology & Molecular Sciences, Johns Hopkins University, Baltimore, MD, March, 2002. 11. Seminar, “Targeting Retroviral RNA: Development of Fluorescence and NMR techniques to identify novel therapeutics” Department of Chemistry, Swarthmore College, Swarthmore, PA, March, 2002. 12. Seminar, “RNA Loop-Loop Complexes, Structural Variations on an Antisense Theme” Annual Meeting of the Biophysical Society, San Francisco, CA, February, 2002. 13. Poster Presentation, M. R. Mihailescu, E. S. DeJong & J. P. Marino “Heteronuclear NMR Studies of the Dimer Initiation Site of HIV-1” International Conference on Magnetic Resonance in Biological Systems, Toronto, CA, August, 2002. 14. Poster Presentation, E.S. DeJong & J. P. Marino “Heteronuclear NMR Studies of the Dimer Initiation Site of HIV-1” International Conference on Magnetic Resonance in Biological Systems, Toronto, Canada, August, 2002. 15. Poster Presentation, E. S. DeJong & J. P. Marino “Proflavin Binds and Stabilizes the Structure of the High-Affinity Rev Binding Site of the Rev Responsive Element of HIV-1 as a Dimer” Annual Meeting of the RNA Society, Madison, WI, May, 2002. 16. Poster Presentation, Luy, B., U. Richter, E. S. DeJong, O.W. Sørensen and J. P. Marino “Observation of H-bond Mediated 3hJH2H3 Couplings across Watson-Crick AU Base Pairs in RNA” Experimental NMR Conference, Monterey, CA, April, 2002. 17. Poster Presentation, B. Luy & J. P. Marino “JE-TROSY: Combined J- and TROSY-Spectroscopy for the Measurement of One-Bond Couplings in Large Macromolecules” European Experimental NMR Conference, Prague, Czechoslovakia, April, 2002. 2001 1. Seminar, “Biomolecular NMR” CARB/NIST Symposium, Rockville, MD, November, 2001.

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2. Seminar, “Targeting RNA: Development of Fluorescence and NMR Techniques to Identify Novel Therapeutics” Department of Medicinal Chemistry, Virginia Commonwealth University, Richmond, VA, October, 2001. 3. Seminar, "Mechanism of NCp7 Mediated HIV-1 DIS Maturation" NIH RNA Interest Group, Bethesda, MD, July, 2001. 4. Seminar, "Targeting Retroviral RNA: Development of Fluorescence and NMR Techniques to Identify Novel Therapeutics" AIDS Structural Biology Meeting, Bethesda, MD, June, 2001. 5. Seminar, “RNA Loop-loop Complexes: Structural Variations on an Antisense Theme” ACS MidAtlantic Regional Meeting, Towson, MD, May, 2001. 6. Seminar, "Targeting RNA: The Development of High-Throughput Screens to Identify Novel therapeutics" NIST Measurements and Materials at the Bio-Interface, Gaithersburg, MD, February, 2001. 7. Poster Presentation, DeJong, E.S. & J.P. Marino "NMR Studies of ColE1 RNA Kissing Hairpin Complex Recognition by the Rom Protein" RNA Society Meeting, Banff, Canada, May, 2001. 8. Poster Presentation, Rist, M. and J.P. Marino “Quantitative analysis of antisense RNA loop-loop complexes using 2-aminopurine fluorescence” Annual Meeting of the Biophysical Society, Boston, MA, February, 2001. 9. Poster Presentation, Luy, B., M. Werner and J.P. Marino “F-19 NMR as a method for monitoring ligand binding to RNA and RNA-protein complexes” Annual Meeting of the Biophysical Society, Boston, MA, February, 2001. 10. Poster Presentation, Marino, J.P., E.S. DeJong, K. Lacourciere, B. Luy, N. Mahashetty and H. Shimada “Structure and Dynamics of the R1 plasmid CopA-CopT RNA Complex” Keystone Symposium on Frontiers of NMR in Molecular Biology V, Big Sky, MT, January, 2001. 2000 1. Seminar, “Structural and Kinetic Studies of RNA Loop-Loop Complexes” Department of Biochemistry, University of Montana, Missoula, MT, November, 2000. 2. Seminar, “Designing and Screening Small Molecules that Target RNA using Fluorescence and NMR Spectroscopy” CHI Meeting on NMR: Drug Discovery and Design, McLean, VA, October, 2000. 3. Seminar, “Structure and Dynamics of the CopA-CopT RNA Complex” Annual Meeting of the RNA Society, Madison, WI, June, 2000. 4. Poster Presentation, Luy, B. and J.P. Marino “Measurement of 19F-1H and 1H-1H Dipolar Coupling Interactions in RNA Oligonucleotides Oriented Using Filamentous Bacteriophage” Experimental NMR Conference, Asilomar, CA, March, 2000.

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5. Poster Presentation, Luy, B. and J.P. Marino “Observation of Scalar Couplings across RNA Base Pair Hydrogen Bonds Mediated by Exchange Broadened Protons” Experimental NMR Conference, Asilomar, CA, March, 2000. 1999 1. Seminar, “NMR Studies of the CopA-CopT RNA Complex” Washington Area NMR Group, NIH, Bethesda, MD, December, 1999. 2. Seminar, “NIST, NMR and the New Millennium” First Joint Biotechnology/Analytical Division Seminar, NIST, Gaithersburg, MD, November, 1999. 3. Seminar, “Fluorescence and NMR Methods for Studying Ligand Binding to RNA” Message Pharmaceuticals Inc., Philadelphia, PA, September, 1999. 4. Poster Presentation, Lacourciere, K.A., Stivers, J.T. and J.P. Marino “Screening Ligand Binding to RNA using Fluorescence and NMR Spectroscopy” CHI Meeting on NMR and Drug Discovery, Baltimore, MD, October, 1999. 5. Poster Presentation, Lacourciere, K.A. and J.P. Marino “Studies of the ColE1 RNA Loop-Loop Kissing Complex Recognition by the Rom Protein” Keystone Symposium on Frontiers of NMR in Molecular Biology IV, Breckenridge, CO, February, 1999. 6. Poster Presentation, Shimada, H., Lacourciere, K.A. and J.P. Marino “Heteronuclear NMR Characterization of the Structural and Dynamics of the CopA-CopT Bimolecular RNA Hairpin Complex” Keystone Symposium on NMR of Macromolecules, Breckenridge, CO, February, 1999. 1998 1. Seminar, “Using Multiple Quantum Coherence to Dramatically Enhancements of Proton-Carbon Correlations in Uniformly 15N,13C-Labeled RNA” Department of Chemistry, California Institute of Technology, Pasadena, CA, May, 1998. 2. Seminar, “NMR Characterization of the Structure and Dynamics of RNA ‘Kissing’ Complexes” NIH RNA Interest Group, Bethesda, MD, June, 1998. 3. Poster Presentation, Marino, J.P., Diener, J., Moore, P.B. and Griesinger, C. “Using Multiple Quantum Coherence to Dramatically Enhancements of Proton-Carbon Correlations in Uniformly 15 13 N, C-Labeled RNA” Experimental NMR Conference, Asilomar, CA, March 1998. 1997 1. Seminar, “Using Multiple Quantum Coherence to Dramatically Enhancements of Proton-Carbon Correlations in Uniformly 15N,13C-Labeled RNA” Department of Chemistry, University of Maryland College Park, College Park, MD, November, 1997. 2. Seminar, “Structure and Stability of RNA Loop-Loop ‘Kissing’ Complexes” Molecular and Cell Biology Program, University of Maryland College Park, College Park, MD, November, 1997.

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1996 1. Seminar, “NMR Characterization of the Structure and Dynamics of an RNA ‘Kissing’ Complex” Department of Biochemistry & Biophysics, University of Pennsylvania, Philadelphia, PA, December, 1996. 2. Seminar, “NMR Characterization of the Structure and Dynamics of an RNA ‘Kissing’ Complex” Department of Biological Chemistry, Harvard Medical School, Boston, MA, December 1996. 3. Seminar, “NMR Characterization of the Structure and Dynamics of an RNA ‘Kissing’ Complex” FASEB Summer Conference, Copper Mountain, CO, June, 1996. 4. Seminar, “Using Multiple Quantum Coherence to Dramatically Enhancements of Proton-Carbon Correlations in Uniformly 15N,13C-Labeled RNA” Department of Chemistry, Frieddrich-Schiller Universität, Jena, Germany, May, 1996. 5. Seminar, “Using Multiple Quantum Coherence to Dramatically Enhancements of Proton-Carbon Correlations in Uniformly 15N,13C-Labeled RNA” Stable Isotope Aided NMR Conference, Frankfurt, Germany, March 1996.

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