Essai de mise en évidence d'un rôle régulateur des tRNA dans la synthèse des protéines de l'œuf de poule

Illinger, D.; Meur, Le; Gerlinger, P; Jp, Ebel

doi:10.1016/s0300-9084(74)80069-6

Cited by 8 publications

(12 citation statements)

References 22 publications

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“…These spots were located in areas indicated on the schematic diagram (Fig. 2) (38), corresponds to initiator tRNAM t. The number of isoaccepting species separated by 2D electrophoresis is in general agreement with that found by fractionation on the RPC 5 columns, as it could be roughly estimated according to chromatographic profiles given in various reports (5,13,34). Two exceptions were observed: tRNAAIa and tRNAMet, which are resolved in two and three spots, respectively, by 2D electrophoresis and in three and four peaks on RPC 5 columns (13, 38).…”

Section: Resultssupporting

confidence: 86%

“…The fact that the tRNA population changes from one tissue to another, as shown by Illinger et al (13) with the livers and oviducts from laying hens, may also be regarded as indicative of adaptation of the tRNA sets in differentiated cells. The adaptation of tRNA contents to the needs of protein synthesis requires a finely regulated mechanism of adjusted tRNA transcription or processing.…”

Section: Resultsmentioning

confidence: 94%

“…Crude aminoacyl-tRNA ligase was prepared from chicken liver by the method of Illinger et al (13 Identification of the fractionated tRNAs. Two methods were used to establish the CEF tRNA gel map by the acceptor specificity of the fractionated tRNA species.…”

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confidence: 99%

“…RNA extracted from the gels was dissolved in a small volume of water (about 120 ,ul for the first-dimension gel bands and 25 ,ul for the second-dimension gel spots). For aminoacylation, the reaction mixture (25 ,ul) according to the optimized values as given by Illinger et al (13) for chicken tRNA. Incubation was at 37°C for 30 min.…”

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confidence: 99%

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Polyacrylamide gel mapping of chicken tRNA: comparison of polysome-bound and whole-cell tRNA from normal and avian sarcoma virus-infected chicken embryo fibroblasts.

Reinisch¹,

Heyman²

1982

Mol. Cell. Biol.

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Analysis of the tRNA population from chicken cells was performed by means of polyacrylamide gel mapping. About 60 species were detected; most of these were positively identified by their acceptor specificity. The comparison of polysomebound and overall cellular tRNA gel patterns from normal and Rous sarcoma virus-infected chicken embryo fibroblasts led us to the following observations: some tRNA species were present in the same relative proportions in all the preparations, and within isoaccepting groups the same species was preponderant; however, although about 8% of whole-cell tRNA was recovered in polysomal preparations, amounts ranging from 3 to 30% were found for individual tRNA species. This points to the absence of a direct correlation between the amount of each mature tRNA species produced and the frequency with which it is used in this case of embryonic cells. No significant difference was observed between the whole-cell tRNA patterns from normal and infected cells. Thus, tRNA transcription appears unaltered when cells are transformed and virus producing. No change was observed in the extent of a post-transcriptional modification of tRNAPhe (the base Y). However, viral infection led to some changes in the relative proportions of individual species from polysomal preparations.Modifications of tRNA population in the cell induced by changes in its environment have been described in numerous reports (23,33). These studies have contributed to opening the question of the role of tRNA in the regulation of translation. Particular attention has been paid to the altered tRNAs in the process of cell differentiation and viral infection. In this last case, some variations in isoacceptor pattern were detected by changes in the chromatographic profiles of amino acid acceptor activity (5,14,31,34 variation in the frequency of codons being translated can occur, involving a concomitant change in the set of tRNAs selected by the polysomal machinery. A strict adjustment of the tRNA population to the amino acid composition of proteins being synthesized has been described for different tissues (6). In this case, the distribution of mature tRNAs in the cell would be the same as the distribution of tRNAs engaged in protein synthesis. We attempted to determine whether virusinduced transformation involves changes either in the amount of each mature tRNA species produced or in the frequency of their utilization or in both. To this end, we compared the distribution of overall cellular tRNAs and that of tRNAs bound to polysomes, assuming that those tRNAs are actively engaged in protein synthesis. Preparations of tRNA isolated from normal CEF and from RSV-infected CEF were analyzed by two-dimensional (2D) gel electrophoresis. The tRNA spots on the gel map were identified. To quantify each species, tRNA was labeled at the 3' end group with [a-32P]

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Section: Resultssupporting

confidence: 86%

Section: Resultsmentioning

confidence: 94%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Polyacrylamide gel mapping of chicken tRNA: comparison of polysome-bound and whole-cell tRNA from normal and avian sarcoma virus-infected chicken embryo fibroblasts.

Reinisch¹,

Heyman²

1982

Mol. Cell. Biol.

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“…Palmiter [3] has shown an increase of the initiation and elongation rates of ovalbumin synthesis under estrogen stimulation which suggests that hormonal control occurs not only at transcription but also at the level of the translation of messenger RNAs into proteins. Our previous work [4,5] indicated that there is a low probability of regulation by the aminoacyl-tRNA synthetases. For example, the seryl-tRNA synthetases prepared either from laying hen liver (estrogen stimulation) or from rooster liver (control) showed the same chromatographic and electrophoretic properties.…”

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Messenger RNA Translation in the Presence of Homologous and Heterologous tRNA

Meur¹,

Gerlinger²,

Ebel³

1976

European Journal of Biochemistry

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The effect of tRNA distribution in the cell on the rate of specific protein synthesis has been investigated. A tRNA-dependent cell-free protein-synthesizing system derived from Krebs-11 ascites cells has been worked out. In this system it is possible to synthesize specific proteins (egg-white proteins or globin) by the translation of oviduct or reticulocyte messenger RNA in the presence of tRNA from homologous or heterologous tissues. The rate of translation of a given messenger RNA is optimal in the presence of tRNA from the homologous tissue. The lower level of protein synthesis obtained with an excess of heterologous tRNA can be overcome by adding the homologous tRNA. Nevertheless homologous isoaccepting species partly purified by a reversed-phase chromatography or a benzoylated DEAE-cellulose column cannot fully restore the optimal synthesis, eliminating the hypothesis according to which a particular tRNA species is involved in this phenomenon. The correct distribution of tRNA is necessary for optimal translation of a messenger RNA.In birds estrogens control the synthesis of eggwhite and yolk proteins [1,2]. Palmiter [3] has shown an increase of the initiation and elongation rates of ovalbumin synthesis under estrogen stimulation which suggests that hormonal control occurs not only at transcription but also at the level of the translation of messenger RNAs into proteins. Our previous work [4,5] indicated that there is a low probability of regulation by the aminoacyl-tRNA synthetases. For example, the seryl-tRNA synthetases prepared either from laying hen liver (estrogen stimulation) or from rooster liver (control) showed the same chromatographic and electrophoretic properties. This enzyme also had the same kinetic constants for the four serylisoaccepting tRNAs either from hens or from other animals such as rabbits or rats. Nevertheless, we have found significant quantitative differences in the distribution of the tRNA isoaccepting species from liver and oviduct (two target organs for estrogens in hens). This observation suggests that a functional adaptation of the tRNA population might exist in response to the requirements of the proteins synthesized in the cell. In order to verify if the tRNA distribution has an influence on the protein synthesis, we developed a tRNA-dependent cell-free system derived from Krebs-I1 ascites cells. In this system we have characterized specific proteins synthesized by translation of Definition. A260 unit, the quantity of material contained in 1 ml of a solution which has an absorbance of 1 at 260 nm, when measured in a I-cm pathlength cell.

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Messenger RNA translation in a tRNA dependent cell‐free system: Role of tRNA from different sources

1975

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Essai de mise en évidence d'un rôle régulateur des tRNA dans la synthèse des protéines de l'œuf de poule

Cited by 8 publications

References 22 publications

Polyacrylamide gel mapping of chicken tRNA: comparison of polysome-bound and whole-cell tRNA from normal and avian sarcoma virus-infected chicken embryo fibroblasts.

Polyacrylamide gel mapping of chicken tRNA: comparison of polysome-bound and whole-cell tRNA from normal and avian sarcoma virus-infected chicken embryo fibroblasts.

Messenger RNA Translation in the Presence of Homologous and Heterologous tRNA

Messenger RNA translation in a tRNA dependent cell‐free system: Role of tRNA from different sources

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