Isolation and partial characterization of Escherichia coli mutants with low levels of transfer ribonucleic acid nucleotidyltransferase

Murray P Deutscher, R. H. Hilderman

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

In order to determine the function of the enzyme transfer ribonucleic acid (tRNA) nucleotidyltransferase in vivo, five mutants of E. coli containing low levels of this enzyme were isolated. Since no selection procedure for such mutants existed, these strains were isolated by assay of large numbers of colonies from a heavily mutagenized stock. A procedure employing cells made permeable to tRNA and ATP was used to screen the large number of colonies required for the isolation. All the mutants contained less than 20% of the normal level of the AMP incorporating activity of tRNA nucleotidyltransferase in extracts prepared by several methods, and the best mutant contained only about 2% of this activity. Three of the mutants also had equally low levels of the cytidine 5' monophosphate incorporating activity of the enzyme. Despite these low activities, the mutant strains displayed relatively normal growth characteristics at all temperatures examined. The enzyme in the mutant strains was not temperature sensitive, nor were any other abnormal biochemical properties detected. tRNA isolated from the mutant strains was missing significant amounts of its 3' terminal adenosine 5' monophosphate residue, amounting to 10 to 15% in the best mutant. However, only small amounts of the terminal cytidine 5' monophosphate residue were missing. The results indicate that tRNA nucleotidyltransferase is involved in some aspect of synthesis or repair of the 3' terminus of tRNA, and that the enzyme is present in large excess over its requirements for this function.

Original languageEnglish
Pages (from-to)621-627
Number of pages7
JournalJournal of Bacteriology
Volume118
Issue number2
StatePublished - Dec 1 1974
Externally publishedYes

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Nucleotidyltransferases
Transfer RNA
Escherichia coli
Cytidine Monophosphate
Enzymes
Adenosine Monophosphate
Temperature
Adenosine Triphosphate
Growth

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Isolation and partial characterization of Escherichia coli mutants with low levels of transfer ribonucleic acid nucleotidyltransferase. / Deutscher, Murray P; Hilderman, R. H.

In: Journal of Bacteriology, Vol. 118, No. 2, 01.12.1974, p. 621-627.

Research output: Contribution to journalArticle

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