RNase PH is essential for tRNA processing and viability in RNase-deficient Escherichia coli cells

Karen Ost Kelly, Nina Bacher Reuven, Zhongwei Li, Murray P Deutscher

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

RNase PH is a Pi-dependent exoribonuclease that can act at the 3′ terminus of tRNA precursors in vitro. To obtain information about the function of this enzyme in vivo, the Escherichia coli rph gene encoding RNase PH was interrupted with either a kanamycin resistance or a chloramphenicol resistance cassette and transferred to the chromosome of a variety of RNase-resistant strains. Inactivation of the chromosomal copy of rph eliminated RNase PH activity from extracts and also slowed the growth of many of the strains, particularly ones that already were deficient in RNase T or polynucleotide phosphorylase. Introduction of the rph mutation into a strain already lacking RNases I, II, D, BN, and T resulted in in viability. The rph mutation also had dramatic effects on tRNA metabolism. Using an in vivo suppressor assay we found that elimination of RNase PH greatly decreased the level of su3+ activity in cells deficient in certain of the other RNases. Moreover, in an in vitro tRNA processing system the defect caused by elimination of RNase PH was shown to be the accumulation of a precursor that contained 4-6 additional 3′ nucleotides following the -CCA sequence. These data indicate that RNase PH can be an essential enzyme for the processing of tRNA precursors.

Original languageEnglish
Pages (from-to)16015-16018
Number of pages4
JournalJournal of Biological Chemistry
Volume267
Issue number23
StatePublished - Aug 15 1992
Externally publishedYes

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Ribonucleases
Transfer RNA
Escherichia coli
Processing
RNA Precursors
Exoribonucleases
Polyribonucleotide Nucleotidyltransferase
Chloramphenicol Resistance
Kanamycin Resistance
Pancreatic Ribonuclease
Mutation
Gene encoding
Kanamycin
Chloramphenicol
Enzymes
Chromosomes
Metabolism
ribonuclease PH
Assays
Nucleotides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Kelly, K. O., Reuven, N. B., Li, Z., & Deutscher, M. P. (1992). RNase PH is essential for tRNA processing and viability in RNase-deficient Escherichia coli cells. Journal of Biological Chemistry, 267(23), 16015-16018.

RNase PH is essential for tRNA processing and viability in RNase-deficient Escherichia coli cells. / Kelly, Karen Ost; Reuven, Nina Bacher; Li, Zhongwei; Deutscher, Murray P.

In: Journal of Biological Chemistry, Vol. 267, No. 23, 15.08.1992, p. 16015-16018.

Research output: Contribution to journalArticle

Kelly, KO, Reuven, NB, Li, Z & Deutscher, MP 1992, 'RNase PH is essential for tRNA processing and viability in RNase-deficient Escherichia coli cells', Journal of Biological Chemistry, vol. 267, no. 23, pp. 16015-16018.
Kelly, Karen Ost ; Reuven, Nina Bacher ; Li, Zhongwei ; Deutscher, Murray P. / RNase PH is essential for tRNA processing and viability in RNase-deficient Escherichia coli cells. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 23. pp. 16015-16018.
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