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Background

Background. NASA experimented with High Magnetic Fields to simulate zero gravity for experiments on plantsPaul and coworkers found that the magnetic field itself, not just the lack of gravity, was causing stress on the plants. 18.9 T. Control. Paul 2005. . Hypothesis. Strong magnetic fields generate subtle perturbations of biomolecules due to the structural diamagnetic anisotropy of the molecules, causing a disruption of normal biochemical function.

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Background

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    2. Background NASA experimented with High Magnetic Fields to simulate zero gravity for experiments on plants Paul and coworkers found that the magnetic field itself, not just the lack of gravity, was causing stress on the plants

    3. Hypothesis Strong magnetic fields generate subtle perturbations of biomolecules due to the structural diamagnetic anisotropy of the molecules, causing a disruption of normal biochemical function

    4. Transcription

    5. T7 RNA Polymerase: The Hand Model

    6. Top View

    7. 1D Model – No Field

    8. 1D Model – Field

    9. Energy Analysis

    10. Force Analysis

    11. Solve for Magnetic Force

    12. Is This Force Reasonable? Other molecular forces creating extreme structural alterations: ~10-10-10-11 N Overstretching DNA Unfolding Titin (muscle) Unfolding DNA hairpin Preventing T7 RNAP from proceeding along the DNA during transcription

    13. Conclusions Experimentally, Some delay in transcript production is indicated at 9 Tesla for T7 RNA polymerase A reduction in transcript production was observed for SP6 RNA polymerase at 9 Tesla Theoretically, a more accurate model is needed Improve k approximations Improve force estimates Include other possible deformations Next: Further test the hypothesis Analyze experiments at 20 and 25 Tesla

    15. Force vs. Field Strength

    16. The Thumb

    17. References Cheetham, Graham M. T., David Jeruzalmi, and Thomas A Steitz (1999) Structural basis for initiation of transcription from an RNA polymerase-promoter complex. Nature (399) 80-83. Gopal, Vijaya et al (1999) Characterization of Structural Features Important for T7 RNAP Elongation Complex Stability Reveals Competing Complex Conformations and a Role for the Non-template strand in RNA Displacement. J. Mol. Biol. (290) 411-431 Lu, Hui and Klaus Schulten (1999) Steered Molecular Dynamics Simulations of Force-Induced Protein Domain Unfolding. PROTEINS: Structure, Function, and Genetics (35) 453-463 Paul, A.-L., R.J. Ferl, B. Klingenberg, J.S. Brooks, A.N. Morgan, J. Yowtak, and M.W. Meisel (2005) Strong Magnetic Field Induced Changes of Gene Expression in Arabidopsis. Materials Processing in Magnetic Fields: Proceedings of the International Workshop on Materials Analysis and Processes in Magnetic Fields (NHMFL, Tallahassee, 17-19 March 2004). To appear fall 2005. Pauling, Linus (1979) Diamagnetic anisotropy of the peptide group. Biophysics (76) 2293-2294. Sousa, Rui, John Rose and B. C. Wang (1994) The Thumb’s Knuckle: Flexibility in the Thumb Subdomain of T7 RNA Polymerase is Revealed by the Structure of a Chimeric T7/T3 RNA Polymerase. Jol. Mol. Biol. (244) 6-12. T7 RiboMax Express Large Scale RNA Production System Technical Bulletin. Promega. (www.promega.com) Tahirov, Tahir H et al. (2002) Structure of a T7 RNA polymerase elongation complex at 2.9 A resolution. Nature (420) 43-50. Wadelton, Kim et al. (2005) Diamagnetic Anisotropy of T7 RNA Polymerase Report of research preformed Summer 2005 as part of NHMFL REU Program. Worchester, D.L. (1978) Structural Origins of diamagnetic anisotropy. Pro.Natl. Acad. Sci. (75) 5475-5477. Worczak, Marianna et al. (2005) Effects of high magnetic fields on in vitro transcription Report of research preformed Summer 2005 as part of NHMFL REU Program.

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