A New Chromatographic System for Increased Resolution of Transfer Ribonucleic Acids<sup>*</sup>

Weiss, Joseph F.; Kelmers, A.D.

doi:10.1021/bi00860a030

Cited by 200 publications

(52 citation statements)

References 27 publications

(14 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The percentages are averages of 46 experiments for the six amino acids. Leucine, lysine, proline, threonine, methionine, serine, phenylalanine, and tyrosine were reacted separately with tRNA, and their isoaccepting species of tRNA were fractionated by reverse phase column chromatography (35). Cochromatography of two populations, representing the different tissues under study, of aminoacyl-tRNA was performed, one being charged with 3H-amino acid and the other with "4C-amino acid.…”

Section: Resultsmentioning

confidence: 99%

The Fractionation of Transfer Ribonucleic Acid from Roots of Pea Seedlings

Vanderhoef

Key

1970

Plant Physiol.

View full text Add to dashboard Cite

Isoaccepting transfer RNA species for several amino acids were fractionated by reverse phase column chromatography. Transfer RNA from dividing cells of pea (Pisum sat ivum) root was compared to that from nondividing cells, and no relative quantitative or qualitative differences were noted for the transfer RNA species for leucine, lysine, proline, threonine, methionine, serine, and phenylalanine. However, certain artifactual differences for serine and phenylalanine were noted. Quantitative differences were observed in tyrosyl-transfer RNA's. Ribonuclease action on tRNA did not contribute to the tRNA species observed.Transfer RNA is involved in the proper translation of the sequence of bases in mRNA into the prescribed order of amino acids in protein (24). Aminoacyl-tRNA4 is central in theories of translational regulation of enzyme synthesis (1,25,27); it is necessary for the end product repression of the enzymes of the amino acid biosynthetic pathways (12,25,26,37) and for the stringent control of RNA synthesis (10,11,23,38).Isoaccepting tRNAs (different tRNA's which accept the same amino acid) have been demonstrated in various organisms for several amino acids. This has been shown in some cases (e.g., Neurospora, rat liver, and Tetrahymena), to be partly due to mitochondria which have their own tRNA's (4,5,8,30). In other cases, there are changes in tRNA populations which correlate with changes in metabolism or development (3,13,17,20,21,28). Infection by phage (14,16,18,33,34,36) and by Herpes virus (29) also leads to changes in the tRNA population in the host cells.There have been three reports of isoaccepting species of tRNA in higher plants. Vold and Sypherd (32) soybean seedlings, but found that only four of these could be charged by synthetases extracted from the hypocotyl, whereas all six were charged by synthetases extracted from cotyledons. Furthermore, the two hypocotyl tRNA's which were not charged by hypocotyl synthetases were present in relatively smaller amounts in the hypocotyl than in the cotyledon. Williams (37) studied in detail the formation of leucyl-tRNA, and demonstrated at least four leucine isoacceptor tRNA's in bean.This is a study of isoaccepting tRNA species for eight amino acids from pea root. Transfer RNA populations of dividing cells are compared to those of nondividing cells. MATERIALS AND METHODSPlant Material. Alaska peas (Pisum sativum, var. Alaska) were germinated in moist absorbent paper for 4 days at about 25 C. The roots were either excised for immediate synthetase or tRNA extraction, or were excised onto Dry Ice and stored at -20 C for further extraction.Extraction and Purification of tRNA. The roots were divided into three sections (7) for the extraction of tRNA: the apical 2 mm (meristematic tissue), the next 5 mm (elongation region), and the remaining approximately 2 cm of root (maturing region). The elongating tissue was usually discarded, and total RNA was extracted (19) from the rapidly dividing cells of the meristem and from the nondividing, fully elongate...

show abstract

Section: Resultsmentioning

confidence: 99%

The Fractionation of Transfer Ribonucleic Acid from Roots of Pea Seedlings

Vanderhoef

Key

1970

Plant Physiol.

View full text Add to dashboard Cite

show abstract

“…[6][7][8][9][10][11][12][13][14] In addition, changes have been reported in the tRNA patterns of differentiating and of neoplastic cells15-20 as well as in cells dividing under different growth conditions. 21' 22 It has not been established, however, whether or not the rate of protein synthesis actually is influenced by variations in the level of tRNA species in the cell.…”

mentioning

confidence: 99%

The Effect of Trna Concentration on the Rate of Protein Synthesis

Anderson¹

1969

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Abstract.-Two in vitro protein-synthesizing systems derived from E. coli have been utilized to demonstrate that the concentration of a tRNA species can regulate the rate of translation of a messenger RNA. (a) The rate of poly-U-directed C'4-phenylalanine incorporation into protein is stimulated by concentrations of tRNA"56 from 1.5 X 10-8 M to 3.0 X 10-6 M, the latter representing a tRNAPhe/70S ribosome ratio of 7. (b) Components of poly U system: Ribosomes from log phase E. coli W3100 were stirred for 18 hr in 0.01 M MgC12, 0.5 M NH4Cl, 0.01 M Tris-Cl, pH 7.6; they were then resuspended and centrifuged twice more in the same buffer.24 The final pellet was dialyzed 566

show abstract

“…Impurities were removed by washing the column with 0.2 M NaCl (in tris-Mg-DDT), and the tRNA was eluted with 1 M NaCl in this buffer. After ethanol precipitation, the tRNA was dissolved in tris-Mg-DDT to a concentration of 5 mg/ml; insoluble material was removed by Abbreviations: tris-Mg-DDT: tris Cl (10 mM, pH 7.4), MgCI2 (10 mM (29). Columns (0.6 x 120 cm) were eluted with 600-ml linear gradients (0.3-0.7 M) of sodium chloride in 20 mm sodium acetate (pH 5.0), 10 mM MgCl2, 1 mm EDTA, and 0.1 mM dithiothreitol at 20 ml/hr.…”

Section: Methodsmentioning

confidence: 99%

Specific Degradation of a Plant Leucyl Transfer Ribonucleic Acid by a Factor in the Homologous Synthetase Preparation

Babcock¹,

Morris²

1973

Plant Physiol.

View full text Add to dashboard Cite

Partially purified aminoacyl synthetase preparations from pea roots (Pisum sativum L. var. Alaska) contain a heat-labile factor which can degrade leucyl-tRNA61eU to a new species. The singular electrophoretic and chromatographic mobilities, the isoprenoid nucleoside content, and the charging characteristics of the new species (designated leucyl-tRNAL'eU), suggest that it is a fragment of tRNA,IeU containing at least that portion of the original molecule extending from the 3' terminus to the anticodon. Conversion appears to be highly specific since neither bulk tRNA, the other leucine tRNA subspecies, nor tyrosine, phenylalanine, or tryptophan tRNAs are susceptible to degradation during incubation with the synthetase preparation.Differentiation or dedifferentiation of organs or cell lines, nutritional stress, and exposure to pathogens are accompanied in many instances by changes in the chromatographic profiles of isoacceptor tRNA species (5,11,21,26,28 Transfer RNA. The method of preparation of tRNA was similar to that used by others for pea (8,27) and soybean tissue (2). Roots, collected in liquid nitrogen, were powdered and then homogenized in 3 volumes of tris-Mg-DDTF buffer containing 0.5% (w/v) sodium dodecyl sulfate. After shaking twice with 0.5 volume of redistilled phenol saturated with tris-Mg-DDT, total RNA was precipitated from the aqueous phase by the addition of 0.1 volume of 1 M potassium acetate (pH 4.5) followed by 2 volumes of 95% ethanol cooled to -20 C. Transfer RNA was extracted from the precipitate by homogenization with 2 volumes of 1 M NaCl containing 0.1 mM dithiothreitol. A further precipitation with ethanol was followed by a brief treatment with DNase (15 min, 4 C, 50 ,ug/ml DNase) after which the solution was deproteinized with phenol and applied to DEAE-cellulose. Impurities were removed by washing the column with 0.2 M NaCl (in tris-Mg-DDT), and the tRNA was eluted with 1 M NaCl in this buffer. After ethanol precipitation, the tRNA was dissolved in tris-Mg-DDT to a concentration of 5 mg/ml; insoluble material was removed by

show abstract

A New Chromatographic System for Increased Resolution of Transfer Ribonucleic Acids^*

Cited by 200 publications

References 27 publications

The Fractionation of Transfer Ribonucleic Acid from Roots of Pea Seedlings

The Fractionation of Transfer Ribonucleic Acid from Roots of Pea Seedlings

The Effect of Trna Concentration on the Rate of Protein Synthesis

Specific Degradation of a Plant Leucyl Transfer Ribonucleic Acid by a Factor in the Homologous Synthetase Preparation

Contact Info

Product

Resources

About

A New Chromatographic System for Increased Resolution of Transfer Ribonucleic Acids*

Cited by 200 publications

References 27 publications

The Fractionation of Transfer Ribonucleic Acid from Roots of Pea Seedlings

The Fractionation of Transfer Ribonucleic Acid from Roots of Pea Seedlings

The Effect of Trna Concentration on the Rate of Protein Synthesis

Specific Degradation of a Plant Leucyl Transfer Ribonucleic Acid by a Factor in the Homologous Synthetase Preparation

Contact Info

Product

Resources

About

A New Chromatographic System for Increased Resolution of Transfer Ribonucleic Acids^*