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Nucleic Acids Research, 1985, Vol. 13, No. 12 4333-4341
© 1985

Articles

Host transfer RNA cleavage and reunion in T4-infected Escherichia coli CTr5x

Gabriel Kaufmann and Michal Amitsur

Biochemistry Department, Tel Aviv University Ramat Aviv, Tel Aviv 69978, Israel

Received April 1, 1985. Revised May 30, 1985. Accepted May 30, 1985.

T4 mutants lacking pol ynucleotlde kinase (pnk– ) or RNA ligase (rli–) do not grow on E. coli CTr5x. During the abortive Infections there accumulate host tRNA fragments that match into two species severed 31 to the anticodon. The CTr5x-specific fragments appear only transiently with wt phage, implicating the affected enzymes in phosphoryl group rearrangement and religatlon [David et al. (1982) Virol. 123, 480]. In a search for the vulnerable host tRNAs and putative relIgatlon products, tRNA ensembles from uninfected E. coli CTr5x or cells infected with various phage strains were fractionated and compared. A tRNA species absent from rli– infected cells but present in uninfected cells or late in wt infection was thus detected. RNase T1 finger prints of this species, isolated before or after wt infection, were compared with that of an in vitro ligated pair of CTr5x-speclfic fragments. The results indicated that this tRNA Is cleaved upon infection and later on restored to it's original or to a very similar form, by polynucleotide kinase and RNA ligase reactions. It is suggested that depletion of such vulnerable host tRNA species underlies the restriction of pnk– or rli– phage on E. coli CTr5x


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