In vitro reconstitution of anticodon nuclease from components encoded by phage T4 and Escherichia coli CTr5X
Amitsur, M.; Morad, I.; Kaufmann, G.
EMBO Journal 8(8): 2411-2415
1989
ISSN/ISBN: 0261-4189 PMID: 2676520 Document Number: 344292
During phage T4 infection of Escherichia coli strains containing the pre locus the host tRNALys 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 polynulceotide 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 tRNALys is driven by an extract from E. coli prrr (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.