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Control of replication of plasmid R1: the intergenic region between copA and repA modulates the level of expression

A Berzal-Herranz1, E G H Wagner1, R Diaz-Orejas1

  • 1Centro de Investigaciones Biologicas (CSIC), Velázquez 144, E-28006 Madrid, Spain.Department of Microbiology. Biomedical Centre, Uppsala University, Box 581, S-751 23 Uppsala, Sweden.

Molecular Microbiology
|August 5, 2017
PubMed

Insights

Mutations in the R1 plasmid intergenic region impact RepA protein synthesis and replication control. These findings highlight the region's importance in regulating plasmid stability and gene expression.

Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • Plasmid R1 replication initiation is rate-limited by the RepA protein.
  • RepA synthesis is primarily regulated by antisense RNA CopA interacting with RepA mRNA leader region CopT.

Purpose of the Study:

  • Characterize mutations in the intergenic region between copA and repA.
  • Investigate the impact of these mutations on RepA expression and plasmid R1 replication control.

Main Methods:

  • Analysis of mini-R1 derivatives with sequence alterations.
  • Assay of repA expression using translational repA-lacZ fusion constructs.
  • Investigation of mutations using copA gene deletion and tac promoter replacement.

Main Results:

  • Most mutations decreased mini-R1 derivative stability and lowered repA expression.
  • Mutations were shown to directly alter repA expression independent of CopA.
  • Specific mutations affected CopA-mediated control by altering RNA-RNA interaction kinetics or copA promoter activity.

Conclusions:

  • The intergenic region between copA and repA is crucial for controlling R1 replication.
  • Mutations in this region can disrupt plasmid stability and gene expression regulation.
  • This region plays a significant role in the intricate control mechanisms of plasmid R1 replication.

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