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In vivo pre-tRNA processing in Saccharomyces cerevisiae.

J P O'Connor1, C L Peebles

  • 1Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260.

Molecular and Cellular Biology
|January 1, 1991
PubMed
Summary

This study reveals the ordered processing of yeast pre-tRNA, with 5' end trimming preceding 3' end trimming. Splicing and end trimming occur independently, not in a fixed sequence.

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Area of Science:

  • Molecular Biology
  • RNA Processing
  • Yeast Genetics

Background:

  • Transfer RNA (tRNA) biogenesis involves intricate processing steps.
  • Intron removal (splicing) and end modifications are crucial for functional tRNA.
  • The precise order of these events in Saccharomyces cerevisiae was not fully elucidated.

Purpose of the Study:

  • To investigate the processing pathways of intron-containing pre-tRNAs in Saccharomyces cerevisiae.
  • To determine the order of splicing and end-trimming reactions.
  • To characterize splicing intermediates and excised intron products.

Main Methods:

  • Filter hybridization using pre-tRNA intron-specific oligonucleotide probes.
  • Classification of RNA species based on size and structure.
  • Direct sequencing of excised intron products.
  • Verification of intermediates using additional specific probes.

Main Results:

  • Detected linear, excised intron products, with one sequenced.
  • Identified various pre-tRNA species differing in 5' leader and 3' extension presence.
  • Confirmed ordered 5' end trimming before 3' end trimming.
  • Observed splicing intermediates and products with unprocessed ends, indicating independent splicing and end-trimming.
  • Deduced a probable order of 3' splice site cleavage before 5' splice site cleavage.

Conclusions:

  • The processing of pre-tRNA in yeast follows an ordered pathway: 5' end trimming precedes 3' end trimming.
  • Splicing and end-trimming are not strictly ordered relative to each other.
  • Splicing can occur either before or after the end-trimming steps.

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