The Role of Individual Exoribonucleases in Processing at the 3′ End of Escherichia coli tRNA Precursors

Zhongwei Li, Murray P Deutscher

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

We have used an in vitro Escherichia coli tRNA processing system to investigate the specific role of individual exoribonucleases in the 3′ maturation of tRNA precursors. The processing of pre-tRNATyrsu+3 and pre-tRNAArg2 was studied using extracts from cells lacking one or multiple exoribonucleases or using purified RNases. Earlier genetic studies had suggested that multiple exoribonucleases contributed to the maturation of tRNA precursors, and this was proven directly in the studies described here. Complete 3′ processing required the combined action of multiple exoribonucleases, and each RNase showed distinct specificities for maturation of the different parts of the 3′ precursor segment. RNase II and polynucleotide phosphorylase were most effective in shortening long 3′ trailer sequences to intermediates with 2-4 extra 3′ residues. Final trimming of the last few 3′ nucleotides of these precursors was carried out most efficiently by RNases T and PH, but the two enzymes differed in their specificity for individual nucleotide positions. Depending on the tRNA precursor, the relative importance of the various RNases to the overall maturation process differed. We also showed that purified exoribonucleases can completely complement mutant extracts and that tRNA maturation can be totally reconstructed in vitro using purified enzymes. These studies provide the first detailed information about the specific role of individual exoribonucleases in tRNA processing, and bring us closer to defining a complete E. coli tRNA maturation pathway.

Original languageEnglish
Pages (from-to)6064-6071
Number of pages8
JournalJournal of Biological Chemistry
Volume269
Issue number8
StatePublished - Feb 25 1994
Externally publishedYes

Fingerprint

Exoribonucleases
RNA Precursors
Escherichia coli
Transfer RNA
Ribonucleases
Processing
Nucleotides
Polyribonucleotide Nucleotidyltransferase
Enzymes
Cell Extracts
Light trailers
Trimming

ASJC Scopus subject areas

  • Biochemistry

Cite this

The Role of Individual Exoribonucleases in Processing at the 3′ End of Escherichia coli tRNA Precursors. / Li, Zhongwei; Deutscher, Murray P.

In: Journal of Biological Chemistry, Vol. 269, No. 8, 25.02.1994, p. 6064-6071.

Research output: Contribution to journalArticle

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