Substrate recognition by RNase P and by the catalytic M1 RNA: identification of possible contact points in pre-tRNAs

EMBO J. 1990 Jun;9(6):1929-37. doi: 10.1002/j.1460-2075.1990.tb08320.x.

Abstract

Modified bases were introduced into pre-tRNAs during in vitro RNA synthesis or by chemical modification. These RNAs were used as substrates for the catalytic M1 RNA and the RNase P holoenzyme from Schizosaccharomyces pombe. The synthetic approach permitted the insertion of 100% m7GTP into pre-tRNAs and this resulted in complete inhibition of the specific 5' processing reactions. Partially modified RNAs were obtained by chemical modifications of purines and uridines in the pre-tRNAs. This allowed detailed analyses of specific bases excluded in the products. With pre-tRNA(Ser) and initiator pre-tRNA(Met), strong effects were observed in the T arm and weaker effects in the anticodon stem. Only minor base exclusions were detected in the acceptor stem of pre-tRNA(Ser) and in the D arm of pre-tRNA(Met).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding Sites
  • Endoribonucleases / metabolism*
  • Models, Molecular
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • RNA Precursors / analysis*
  • RNA Processing, Post-Transcriptional
  • RNA, Transfer / analysis*
  • Ribonuclease P
  • Saccharomycetales / genetics*
  • Schizosaccharomyces / enzymology
  • Schizosaccharomyces / genetics*
  • Substrate Specificity

Substances

  • RNA Precursors
  • RNA, Transfer
  • Endoribonucleases
  • Ribonuclease P