Synthesis of Transfer Ribonucleic Acid in KB Cells Infected with Adenovirus Type 2

Lm, Sehulster; Varricchio, Frederick; Jun, Raska K

doi:10.1099/0022-1317-40-1-183

“…However, the number of tRNA species detected by 2D gel electrophoresis reaches about 60 when taking into account the comigration of some species and the presence of six additional spots identified in the chicken liver tRNA gel map. This number is close to that found for other eucaryotic tRNA preparations resolved by 2D gel electrophoresis (7,19,30). However, more tRNA species than those identified could map in some spots since we cannot exclude a loss of acceptor activity of some tRNAs after gel elution.…”

Section: Resultsmentioning

confidence: 55%

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

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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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“…The acceptance activities of the total tRNA extracted from the identified spots (as given in Table 2) were summarized after deduction of the blank values. oacylation values ranging between 6 and 24 pmol of amino acid per 30 pmol of tRNA were found.…”

Section: Resultsmentioning

confidence: 95%

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

Molecular and Cellular Biology

0

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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 polysome-bound 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.

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“…normal form versus the suppressor form of E. coli tRNATrP (base substitution C -. U in the anticodon [23]), the effect of tRNA modifications on gel migration has not been analyzed in any rigorous manner and can only be inferred from other data (9,11,27,31). We feel that it is likely that a tRNA base modification such as the thiomethylation reported to occur for some tRNA species during sporulation (2,15,21,37) would not produce an altered tRNA migration pattern in the 2-D gel system that we used.…”

Section: Discussionmentioning

confidence: 99%

“…The GSY266 strain of B. subtilis is a metB3 derivative of strain 168M oriinally from the laboratory of C. Anagnostopoulos (38). The sponlation medium (LDSM) and nutrient agar plating medium (NAT) have been described previously (31). Low-phosphate (low-P) LDSM medium was prepared by treating the nutrient broth component of LDSM with NH4OH and MgCl2 as described by Pieczenik et al (28).…”

Section: Methodsmentioning

confidence: 99%

Transfer ribonucleic acid synthesis during sporulation and spore outgrowth in Bacillus subtilis studied by two-dimensional polyacrylamide gel electrophoresis

Henner¹,

Steinberg²

1979

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The synthesis of transfer ribonucleic acid (tRNA) was examined during spore formation and spore outgrowth in Bacillus subtilis by two-dimensional polyacrylamide gel electrophoresis of in vivo 32P-labeled RNA. The two-dimensional gel system separated the B. subtilis tRNA's into 32 well-resolved spots, with the relative abundances ranging from 0.9 to 17% of the total. There were several spots (five to six) resolved which were not quantitated due to their low abundance. All of the tRNA species resolved by this gel system were synthesized at every stage examined, including vegetative growth, different stages of sporulation, and different stages of outgrowth. Quantitation of the separated tRNA's showed that in general the tRNA species were present in approximately the same relative abundances at the different developmental periods. tRNA turnover and comt Present address:

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Synthesis of Transfer Ribonucleic Acid in KB Cells Infected with Adenovirus Type 2

Cited by 7 publications

References 28 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.

Polyacrylamide Gel Mapping of Chicken tRNA: Comparison of Polysome-Bound and Whole-Cell tRNA from Normal and Avian Sarcoma Virus-Infected Chicken Embryo Fibroblasts

Transfer ribonucleic acid synthesis during sporulation and spore outgrowth in Bacillus subtilis studied by two-dimensional polyacrylamide gel electrophoresis

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