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B R Reid

Publications and source records attributed to B R Reid.

107 records · Page 6Linked to original sources
PubMed1973

Determination of secondary and tertiary structural features of transfer RNA molecules in solution by nuclear magnetic resonance.

High-resolution 300-MHz proton nuclear magnetic resonance spectra of the hydrogen-bounded protons in three different purified tRNA molecules are presented. The resonances in the region between -11 and -15 ppm from 2,2-dimethyl-2-silapentane-5-sulfonate (DSS) are assigned to the ring NH protons of specific base pairs by two approaches. First, intrinsic positions of -14.8 ppm and -13.7 ppm are taken for the AU and GC ring NH protons, respectively, and the spectra are calculated by including ring current shifts from the nearest neighbors. The spectra calculated in this way on the basis of the cloverleaf are in good agreement with the observed. Second, fragments of yeast tRNA(Phe) were obtained, which helped in assignments of the spectrum of intact molecules. The close agreement strongly supports the cloverleaf model. Tertiary structural features were determined in a few cases where the ring currents at the terminal base pairs of helical regions depended upon stacking of the helices. In this way, we were able to show that in Escherichia coli tRNA(Glu) the CCA stem forms a continuous helix with the TPsiC stem, which is in accord with the preliminary x-ray structure of yeast tRNA(Phe), suggesting that this stacking is observed in solution and may be a general property of different tRNA molecules. Similar reasoning suggests that in E. coli tRNA(fMet) G-27 is stacked upon the dihydrouridine helix.

Base Sequence↗
PubMed1973

Conformation of charged and uncharged tRNA.

The effect of aminoacylation on the conformation of yeast tRNA(Phe) was investigated by high-resolution (300 MHz) proton nuclear magnetic resonance (NMR) spectroscopy. Resonances in the low-field (-11 to -15 ppm) region of the spectra are due to ring NH protons of Watson-Crick base pairs, and to a very high degree of approximation (within 0.05 ppm) the low-field spectra of tRNA(Phe) and phenylalanyl-tRNA(Phe) are identical. From this observation and analysis of the low-field NMR spectra we conclude that the secondary structures of the two tRNAs are identical with respect to base-pairing schemes and interbase distances in the helical region (0.1-0.2 A). Several tertiary structural features, including conformation of the dihydro-U loop, conformation of the minor loop, relative orientations of the acceptor and the TPsiC stems, dihydro-U and anticodon stems, and probably conformation of the anticodon loop are shown to be the same in tRNA(Phe) and phenylalanyl-tRNA(Phe). Our results leave little remaining opportunity for changes in tertiary structure that would not have been observed by the NMR method.

Base Sequence↗
PubMed1970

Changes in Nucleic Acids in Phytochrome-dependent Elongation of the Alaska Pea Epicotyl.

Red light, which produces the physiologically active form of phytochrome (Pfr), inhibited epicotyl elongation in intact dark-grown Alaska pea seedlings. This red light response was detectable 3 hours after the light treatment and became pronounced after 5 hours. The growth inhibition was completely reversed by far red light applied immediately after the red or by pretreatment of the seedlings with the plant hormone gibberellin A(3).Comparison of the total (32)P-labeled nucleic acids from control and red light-treated Alaska pea epicotyls on methylated albumin-kieselguhr columns revealed a marked alteration of the pattern of nucleic acid synthesis in this plant material with little or no effect on total isotope incorporation into nucleic acids. A single 5-minute red light perturbation caused a 2-fold stimulation of (32)P incorporation into the tRNA fraction while, simultaneously, (32)P incorporation into tenaciously bound RNA was reduced to 50% of control levels. Red light treatment had no effect on (32)P incorporation into the DNA-RNA, rRNA, or mRNA fractions. Far red light reversed the effect of red light on tRNA synthesis but did not restore tenaciously bound RNA levels to the control value. Gibberellin A(3) treatment did not cause reversal of any of the red light effects on RNA synthesis.These light-induced changes in nucleic acids were measurable before any changes in the physiological response (epicotyl elongation) could be detected. These results are consistent with a phytochrome-mediated differential gene activation mechanism in the Alaska pea epicotyl elongation system.

Journal Article↗