Construction and processing of transfer RNA precursor models

Christopher K. Surratt, Zbigniew Lesnikowski, Aria L. Schifman, Francis J. Schmidt, Sidney Hecht

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Several "dimeric" tRNA molecules were constructed as potential substrates for ribonuclease P (RNase P) and for M1 RNA, the catalytic subunit of RNase P. Construction was effected by the T4 RNA ligase-mediated coupling of a mature Escherichia coli tRNA (acceptor substrate) and nucleotides 1-36 of yeast tRNAPhe (donor substrate), followed by annealing of the 3′-half of yeast tRNAPhe (nucleotides 38-76). E. coli RNase P and M1 RNA were both found to cleave the dimeric tRNA precursor model constructed from E. coli tRNAPhe (5′-tRNA) and yeast tRNAPhe (3′-tRNA) in a reaction that was dependent on the presence of the annealed 3′-half molecule derived from yeast tRNAPhe, or on some conformation imposed by the presence of this species; the product had the same mobility as authentic E. coli tRNAPhe on a polyacrylamide gel. By utilizing tRNA precursor models radiolabeled at phosphodiesters immediately preceding or following the putative site of processing, cleavage of the substrate by both M1 RNA and the holoenzyme was demonstrated to occur at the expected phosphate ester linkage. The results obtained here suggest that the endonucleolytic separation of two tRNAs by RNase P is dependent on one or more structural features in the 3′-half of the 3′-tRNA, and thus are consistent with the report of McClain et al. (McClain, W. H., Guerrier-Takada, C., and Altman, S. (1987) Science 238, 527-530) that identifies the T stem and loop as a possible recognition site.

Original languageEnglish (US)
Pages (from-to)22506-22512
Number of pages7
JournalJournal of Biological Chemistry
Volume265
Issue number36
StatePublished - Dec 25 1990
Externally publishedYes

Fingerprint

RNA, Transfer, Phe
RNA Precursors
Transfer RNA
Ribonuclease P
Yeast
Processing
Yeasts
Escherichia coli
RNA
Substrates
Nucleotides
Catalytic RNA
Holoenzymes
Molecules
Ligases
Conformations
Esters
Phosphates
Annealing

ASJC Scopus subject areas

  • Biochemistry

Cite this

Surratt, C. K., Lesnikowski, Z., Schifman, A. L., Schmidt, F. J., & Hecht, S. (1990). Construction and processing of transfer RNA precursor models. Journal of Biological Chemistry, 265(36), 22506-22512.

Construction and processing of transfer RNA precursor models. / Surratt, Christopher K.; Lesnikowski, Zbigniew; Schifman, Aria L.; Schmidt, Francis J.; Hecht, Sidney.

In: Journal of Biological Chemistry, Vol. 265, No. 36, 25.12.1990, p. 22506-22512.

Research output: Contribution to journalArticle

Surratt, CK, Lesnikowski, Z, Schifman, AL, Schmidt, FJ & Hecht, S 1990, 'Construction and processing of transfer RNA precursor models', Journal of Biological Chemistry, vol. 265, no. 36, pp. 22506-22512.
Surratt CK, Lesnikowski Z, Schifman AL, Schmidt FJ, Hecht S. Construction and processing of transfer RNA precursor models. Journal of Biological Chemistry. 1990 Dec 25;265(36):22506-22512.
Surratt, Christopher K. ; Lesnikowski, Zbigniew ; Schifman, Aria L. ; Schmidt, Francis J. ; Hecht, Sidney. / Construction and processing of transfer RNA precursor models. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 36. pp. 22506-22512.
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