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Nucleic Acids Research, 1978, Vol. 5, No. 1 257-269
© 1978

Articles

1H NMR of valine tRNA modified bases. Evidence for multiple conformations

Rodney V. Kastrup+ and Paul G. Schmidt*

School of Chemical Sciences, University of Illinois Urbana, IL 61801, USA

Received November 17, 1977.

Methyl and methylene protons of dihydrouridine 17 (hU), 6-methyladenosine 37 (m6A), 7-methylguanosine 46 (m7G) and ribothymidine 54 (rT) give clearly resolved peaks (220 MHz) for tRNA1val (coil) solutions in D20, 0.25 m NaCl, at 27° C. Chemical shifts are generally consistent with a solution structure of tRNA1val similar to the crystal structure of tRNAphe (yeast). At least 3 separate transitions are observed as the temperature is raised. The earliest involves disruption of native tertiary structure and formation of intermediate structures in the m7G and rT regions. A second transition results in a change in structure of the anticodon loop, containing m6A. The final step involves unfolding of the m7G and rT intermediates and melting of the T{Psi}C helix. Low salt concentrations produce multiple, partially denatured conformations, rather than a unique form, for tRNA1val. Native structure is almost completely reformed by addition of Na+ but Mg2+ is required for correct conformation in the vicinity of m7G.

Present address: Department of Chemistry, University of Rochester, Rochester, New York 14627

Present address: Oklahoma Medical Research Foundation, 825 N. E. 13th St., Oklahoma City, Oklahoma 731014.


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