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• W2090955348 abstract "Tritium exchange was used as a probe of transfer RNA structure in experiments with unfractionated tRNA (tRNAUnfrac and homogeneous tRNA3Leu from bakers' yeast. Exchange kinetics were measured over a range of ionic conditions that vary in ability to stabilize the secondary and tertiary structure of tRNA. The native conformations of both samples show the same kinetics of exchange. The kinetics for tRNA3Leu trapped in a denatured state in a “native” solvent are much faster, reflecting the conformation and not the ionic medium. In 0.1 M-Na+, where tRNA3Leu is denatured, the kinetics for tRNAUnfrac are intermediate between those for native and denatured tRNA3Leu, suggesting that in this solvent at 0 °C some tRNAs are denatured whereas other are still native. Upon further lowering of Na+ concentration, tRNAUnfrac shows increasingly faster exchange, suggesting complete electrostatic denaturation of the tertiary structure of all the tRNAs in the sample, and even disruption of secondary structure. Extrapolation of the essentially linear early-time kinetics to zero time provides minimal estimates of the number of slowly exchanging hydrogens. For native tRNA3Leu the number is 111±2 hydrogens, whereas for the trapped denatured conformation it is only 95±2. This difference reflects a smaller number of hydrogen-bonded bases in the denatured conformation. In 1 M-Na+, 101±2 slowly exchanging hydrogens are found for the native tRNA3Leu conformation, suggesting an incompletely formed native structure. For native tRNAUnfrac the comparable number is 101±3. These numbers of slowly exchanging hydrogens in the native conformations are consistent with tertiary structural hydrogen-bonding. Furthermore, this tertiary structure must be responsible for the slower exchange by native tRNA. The observed numbers of exchangeable hydrogens provide a basis for comparison of hydrogen-bonding interactions in native and denatured tRNA conformations. The mechanism of renaturation was also investigated, using tritium exchange as a monitor of perturbation of base pairing during the transition. When tRNAUnfrac in low Na+ is renatured by addition of Mg2+ during tritium exchangeout, a burst of exchange or “spillage” of tritium is detected. This suggests that a fraction of the base pairs of the rapidly renaturing tRNAs in the mixture is disrupted during renaturation. In that event, and by analogy with tRNA3Leu, part of the base-pairing arrangement of the denatured conformations may not be preserved in the native state; and if the native conformation includes the full “cloverleaf” pattern of secondary structure, that pattern may not be intact in some denatured conformations." @default.
• W2090955348 created "2016-06-24" @default.
• W2090955348 creator A5009996418 @default.
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• W2090955348 date "1973-03-01" @default.
• W2090955348 modified "2023-09-26" @default.
• W2090955348 title "Tritium exchange studies of transfer RNA in native and denatured conformations" @default.
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• W2090955348 doi "https://doi.org/10.1016/0022-2836(73)90379-3" @default.
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