Transfer RNA metabolism in Escherichia coli cells deficient in tRNA nucleotidyltransferase

Murray P Deutscher, Jim Jung Ching Lin, Jeffrey A. Evans

Research output: Contribution to journalArticle

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Abstract

We have investigated the role of tRNA nucleotidyltransferase in transfer RNA metabolism using mutants (cca mutants) of Escherichia coli deficient in this enzyme. Extracts of the most defective strain contained <2% and <1% of the AMP and CMP-incorporating activities, respectively. About 10% of the tRNA from this cell contained incomplete termini, and this level of defective tRNA was not affected by a variety of growth conditions or by transduction of the cca mutation into a variety of genetic backgrounds. However, tRNA from stationary phase cells was somewhat more defective. Aminoacylation studies revealed that most tRNAs were unaffected by the presence of the cca mutation, but a few species were as much as 20 to 60% defective, and this was true for all the isoacceptors for a given amino acid. Incubation of the mutant cells with the antibiotics chloramphenicol and rifampicin, alone or in combination, indicated that most defective tRNA species arose from end-turnover of the terminal AMP residue. However, the possibility exists that tRNACys, the most defective tRNA species, may require tRNA nucleotidyltransferase for its biosynthesis. No evidence was obtained for a role of the enzyme in the biosynthesis of any other tRNA species. End-turnover of terminal residues was shown to be related to the state of charging of a tRNA molecule, but the physiological significance of this process remains to be determined. Possible explanations for the apparently limited role of tRNA nucleotidyltransferase in E. coli are discussed.

Original languageEnglish
Pages (from-to)1081-1094
Number of pages14
JournalJournal of Molecular Biology
Volume117
Issue number4
DOIs
StatePublished - Dec 25 1977
Externally publishedYes

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Transfer RNA
Escherichia coli
Adenosine Monophosphate
RNA, Transfer, Cys
Transfer RNA Aminoacylation
Physiological Phenomena
Aminoacylation
Cytidine Monophosphate
Mutation
tRNA nucleotidyltransferase
Chloramphenicol
Enzymes
Rifampin
Anti-Bacterial Agents
Amino Acids
Growth

ASJC Scopus subject areas

  • Virology

Cite this

Transfer RNA metabolism in Escherichia coli cells deficient in tRNA nucleotidyltransferase. / Deutscher, Murray P; Lin, Jim Jung Ching; Evans, Jeffrey A.

In: Journal of Molecular Biology, Vol. 117, No. 4, 25.12.1977, p. 1081-1094.

Research output: Contribution to journalArticle

Deutscher, Murray P ; Lin, Jim Jung Ching ; Evans, Jeffrey A. / Transfer RNA metabolism in Escherichia coli cells deficient in tRNA nucleotidyltransferase. In: Journal of Molecular Biology. 1977 ; Vol. 117, No. 4. pp. 1081-1094.
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