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NHMFL 2005 Research Report - University of Florida

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									                                      NATIONAL HIGH MAGNETIC FIELD LABORATORY
                                                2005 RESEARCH REPORT

EFFECTS OF HIGH MAGNETIC FIELDS ON IN VITRO TRANSCRIPTION

Marianna Worczak (UF, Physics via NHMFL REU; and Clarkson University, Chemistry), Kimberly
Wadelton (UF, Physics via NHMFL REU; and Sweet Briar College, Physics), James Ch. Davis (UF,
Physics), Anna-Lisa Paul (UF, Horticultural Sciences), and Mark W. Meisel (UF, Physics)


Introduction
The study of biological molecules in high magnetic fields began in the early 1970’s, when major advances in NMR
instrumentation allowed researchers to observe the partial alignment of nucleic acids and certain phospholipids. Magnetic
effects on protein orientation were originally considered in theoretical calculations of the magnetic anisotropy of peptide
bonds [1,2]. The alignment of biological molecules in magnetic fields has raised the question of how biological processes
might be affected by the presence of strong magnetic fields. Work involving exposure of Arabidopsis thaliana to high
magnetic fields noted an induction of a gene reporter driven by the alcohol dehydrogenase promoter [3]. This observation
suggested that the magnetic field impacts normal biological function, which affected gene expression. Since many reactions
and environmental interactions influence gene expression in a living system, our present work investigates in vitro
transcription of a single process.

Experimental Details
Commercially available in vitro transcription kits contain RNA polymerases isolated from bacteriophages. These
polymerases, such as T7 and SP6 as used in our work, are single unit globular proteins, which maintain active site structure
and function similar to multisubunit polymerases typically found in higher organisms. Since phage polymerases have faster
kinetics than their multisubunit homologs, large amounts of RNA can be quickly produced for study. The working
hypothesis fueling our experiments was that strong magnetic fields acting on the magnetic anisotropies already present in the
RNA polymerases would lead to an alteration in the three dimensional shape of the protein. This alteration, whether it is a
partial alignment or substantial distortion, was hypothesized to have some effect on enzymatic function.

Results and Discussion
To test the hypothesis, experiments employing T7 and SP6 polymerase in vitro transcription kits were preformed in the
presence of magnetic fields ranging up to 25 T. Preliminary results analyzed by gel electrophoresis indicate that transcripts
were not truncated. However, T7 results suggest a subtle reduction in efficiency with increasing magnetic field, which
indicates the potential of some minor effects of the magnetic field on transcription. Results from SP6 indicate an overall
decrease in transcript produced for reactions at 9 Tesla but not at 4.5 Tesla. If our results indeed arise from an alteration of
polymerase conformation, the observed difference in effects between SP6 and T7 reactions may be due to their slightly
different structures. Future tests are necessary to verify the effects observed. In addition, experiments in simple organisms
such as bacteria may allow for control of some variables confounding original plant studies, while still providing a system
more sensitive to subtle magnetic field effects than detectable with the in vitro kits.

Acknowledgements
This work was partially supported by the National High Magnetic Field Laboratory (NHMFL) Summer 2005 REU Program
at the University of Florida in Gainesville. Work was also supported in part by NSF DMR-0305371 and NASA grant
NNA04CC61. We acknowledge R.J. Ferl and W.B. Gurley for enlightening conversations.

References
[1] Worcester, D.L., Proc. Nat. Acad. Sci. USA, 75, 5475-5477 (1978).
[2] Pauling, L., Biophysics, 76, 2293-2294 (1979).
[3] Paul, A.-L., et al., in Magnetic Fields: Proceedings of the International Workshop on Materials Analysis and Processing
in Magnetic Fields, edited by Hans J. Schneider-Muntau and Hitoshi Wada (World-Scientific, Singapore, 2005) pp. 238-242.

								
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