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Bacillus subtilis pur operon expression and regulation.

D J Ebbole1, H Zalkin

  • 1Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907.

Journal of Bacteriology
|April 1, 1989
PubMed
Summary

Bacillus subtilis pur operon transcription initiates at a single promoter. Enhanced translational efficiency in purC gene expression suggests post-transcriptional regulation, impacting purine biosynthesis.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Gene Regulation

Background:

  • The Bacillus subtilis pur operon is a complex 12-gene cluster involved in purine biosynthesis.
  • Gene organization includes overlapping coding units and intercistronic gaps, suggesting intricate regulatory mechanisms.

Purpose of the Study:

  • To investigate the transcriptional and translational regulation of the Bacillus subtilis pur operon.
  • To determine the role of intercistronic regions and translational efficiency in gene expression.

Main Methods:

  • Construction of translational fusions using Escherichia coli lacZ with purE, purC, and purM genes.
  • Analysis of gene fusions integrated into the chromosomal pur operon.
  • Enzyme and mRNA measurements under repressing and nonrepressing purine conditions.

Main Results:

  • Transcription initiates from a single promoter at the 5' end of the pur operon, with regulation occurring solely at this site.
  • Expression of purC-lacZ fusion was significantly higher (3.0- to 6.8-fold) than purE-lacZ due to enhanced translational efficiency.
  • Enhanced translational efficiency of purC-lacZ correlated with a partial escape from purine-mediated regulation.

Conclusions:

  • Transcriptional regulation of the Bacillus subtilis pur operon is primarily controlled at its 5' end.
  • Post-transcriptional regulation, specifically enhanced translational efficiency of purC, plays a significant role in gene expression and purine metabolism.

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