In vitro reconstitution of anticodon nuclease from components encoded by phage T4 and Escherichia coli CTr5X

M. Amitsur, I. Morad, G. Kaufmann

Research output: Contribution to journalArticlepeer-review

Abstract

During phage T4 infection of Escherichia coli strains containing the prr locus the host tRNA(Lys) undergoes cleavage-ligation in reactions catalyzed by anticodon nuclease, polynucleotide kinase and RNA ligase. Known genetic determinants of anticodon nuclease are prr, which restricts T4 mutants lacking polynucleotide kinase or RNA ligase, and stp, the T4-suppressor of prr encoded restriction. The present communication describes an in vitro anticodon nuclease assay in which the specific cleavage of tRNA(Lys) is driven by an extract from E. coli prr(r) (restrictive) cells infected by phage T4. The in vitro anticodon nuclease reaction requires factor(s) encoded by prr, is stimulated by a synthetic Stp polypeptide and appears to require additional T4 induced factor(s) distinct from Stp.

Original languageEnglish
Pages (from-to)2411-2415
Number of pages5
JournalEMBO Journal
Volume8
Issue number8
DOIs
StatePublished - 1989

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