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Related Experiment Videos

Drop-off during ribosome hopping.

A J Herr1, N M Wills, C C Nelson

  • 1Department of Human Genetics, University of Utah, 2030 E. 15N., Salt Lake City, UT 84112-5330, USA.

Journal of Molecular Biology
|August 9, 2001
PubMed
Summary
This summary is machine-generated.

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Bacteriophage T4 gene 60 uses ribosome bypassing to synthesize proteins. While most ribosomes initiate bypassing, only half resume translation, with spontaneous peptidyl-tRNA release being a key factor in this inefficiency.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Bacteriophage T4 gene 60 encodes a topoisomerase subunit.
  • Ribosomes translate mRNA, but can bypass specific sequences.
  • Gene 60 mRNA contains two open reading frames (ORFs) separated by a 50-nucleotide non-coding segment.

Purpose of the Study:

  • To investigate the mechanism of ribosome bypassing in bacteriophage T4 gene 60.
  • To identify factors contributing to inefficient translation resumption after bypassing.
  • To elucidate the reasons for unproductive bypassing events.

Main Methods:

  • Analysis of ribosome bypassing efficiency in bacteriophage T4 gene 60.
  • Assessment of the role of the stop codon in the first ORF.
  • Evaluation of mutant tRNA(Gly)(2) variants' impact on bypassing.

Related Experiment Videos

  • Investigation of factors inducing peptidyl-tRNA drop-off, including ribosome recycling factor.

    • Main Results:

    • Nearly all ribosomes initiate bypassing the 50-nucleotide segment in gene 60 mRNA.
    • Only 50% of ribosomes successfully resume translation in the second ORF.
    • Failure to resume translation is independent of the first ORF's stop codon.
    • Mutant tRNA(Gly)(2) variants decrease bypassing efficiency.
    • Ribosome recycling factor contributes minimally to unproductive bypassing.
    • Spontaneous peptidyl-tRNA release appears to be a major cause of unproductive bypassing.

    Conclusions:

    • Ribosome bypassing in bacteriophage T4 gene 60 is inefficient, with significant unproductive bypassing.
    • Spontaneous peptidyl-tRNA release is likely inherent to the gene 60 bypassing mechanism.
    • This inefficiency may be linked to the specific requirements of synthesizing the topoisomerase subunit.