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Related Experiment Videos

Gene expression in the Streptomyces temperate phage phi C31.

M C Smith1, C J Ingham, C E Owen

  • 1Department of Biological and Molecular Science, University of Stirling, Scotland, UK.

Gene
|June 15, 1992
PubMed
Summary

The Streptomyces phage phi C31 repressor gene produces three proteins from a single locus. The 42-kDa repressor protein likely autoregulates transcription by binding to a conserved DNA sequence.

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Area of Science:

  • Molecular Biology
  • Microbial Genetics
  • Bacteriophage Research

Background:

  • The temperate Streptomyces phage phi C31 repressor gene (c) was previously cloned and sequenced, predicted to encode a single 74-kDa protein.
  • Initial studies suggested a single repressor protein, but further investigation was needed to understand the gene's full expression and regulatory mechanisms.

Purpose of the Study:

  • To elucidate the complete protein products of the Streptomyces phage phi C31 repressor gene (c).
  • To investigate the autoregulation mechanism of the c locus transcription.
  • To identify potential regulatory elements and their distribution within the phi C31 genome.

Main Methods:

  • Gene sequencing and transcript analysis to identify all protein products.
  • Genetic and biochemical assays to probe protein-DNA interactions.

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  • Bioinformatic analysis to identify conserved sequences (inverted repeats) across the phage genome.

    • Main Results:

    • The c gene produces three in-frame proteins (74, 54, and 42 kDa) with identical C-termini but different N-termini, translated from a nest of transcripts.
    • Genetic and biochemical evidence indicates autoregulation of the c locus, likely mediated by the 42-kDa repressor protein.
    • A conserved 16-bp perfect inverted repeat sequence, implicated in autoregulation, is found at multiple loci within the phi C31 genome.

    Conclusions:

    • The Streptomyces phage phi C31 repressor gene (c) exhibits complex expression, yielding multiple functional proteins from a single locus.
    • Autoregulation of the c locus by the 42-kDa repressor protein binding to a conserved inverted repeat is a key regulatory mechanism.
    • The conserved inverted repeat sequence suggests a broader role in phi C31 genome regulation, potentially influencing transcription of other genes.