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Enterococcus faecalis plasmid pAD1 replication and maintenance

¹Department of Microbiology, School of Medicine, University of South Dakota, Vermillion, USA.

Developments in Biological Standardization

|January 1, 1995

Summary

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A newly identified DNA fragment, par, ensures stable inheritance of the E. faecalis plasmid pAD1 by producing two small RNAs. These par RNAs are crucial for plasmid stability and partition function.

Area of Science:

Molecular Biology
Genetics
Microbiology

Background:

The stable inheritance of plasmids is crucial for bacterial populations.
Plasmid pAD1 from E. faecalis is pheromone-responsive and requires specific elements for stable maintenance.

Purpose of the Study:

To identify and characterize the determinant responsible for the stable inheritance of the E. faecalis plasmid pAD1.
To elucidate the role of small RNAs in plasmid stability and segregation.

Main Methods:

Isolation and characterization of the par determinant.
Analysis of par-encoded small RNAs (RNAI and RNAII) using molecular techniques.
Site-directed mutagenesis and transposon mutagenesis to assess RNA function.
Testing the function of the isolated par determinant in a heterologous vector.

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Main Results:

A 457 bp DNA fragment, termed par, was identified as essential for stable plasmid inheritance.
The par determinant directs the production of two small RNAs, RNAI (~250 nt) and RNAII (~145 nt).
Mutations affecting par RNA production correlated with decreased plasmid stability, highlighting the RNAs' central role.
The isolated par determinant successfully stabilized a heterologous vector, indicating its self-sufficiency.

Conclusions:

The par determinant and its small RNAs are sufficient for ensuring stable plasmid inheritance.
The small RNAs likely play a structural role in the plasmid partition apparatus rather than being translated.
This study identifies key elements for understanding plasmid stability mechanisms in E. faeacalis.