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The fate of phage lambda DNA in lambda-infected minicells.

H Witkiewicz, K Taylor

Biochimica Et Biophysica Acta

|August 29, 1979

Summary

Bacteriophage lambda DNA binds to Escherichia coli minicell membranes and forms supercoiled structures. Phage infection halts plasmid DNA replication, with only minimal, non-replicative lambda DNA synthesis observed.

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Area of Science:

Microbiology
Molecular Biology
Virology

Background:

Bacteriophage lambda is a model virus extensively studied in molecular biology.
Escherichia coli minicells are small, anucleate derivatives of E. coli used to study surface phenomena and gene expression.
ColE1 is a well-characterized plasmid in E. coli, essential for understanding plasmid-host interactions.

Purpose of the Study:

To investigate the fate of bacteriophage lambda DNA within Escherichia coli minicells.
To determine the impact of phage lambda infection on ColE1 plasmid DNA synthesis.
To characterize the initial synthesis and structural state of lambda DNA post-infection in minicells.

Main Methods:

Infection of E. coli minicells (both plasmid-free and ColE1-harboring) with bacteriophage lambda.
Analysis of lambda DNA binding to the minicell membrane.
Assessment of DNA synthesis using radiolabeling techniques.
Characterization of DNA structures (e.g., supercoiling).

Main Results:

Demonstrated binding of lambda DNA to the minicell membrane.
Confirmed the formation of supercoiled, covalently-closed circular lambda DNA structures.
Observed a complete abolition of ColE1 plasmid DNA synthesis upon phage infection.
Detected only slight, non-replicative synthesis of lambda DNA fragments shortly after infection.

Conclusions:

Phage lambda DNA interacts with the E. coli minicell membrane, adopting a supercoiled form.
Lambda infection effectively inhibits host plasmid replication.
Early lambda DNA synthesis is limited and associated with fragmented DNA, suggesting a non-replicative process.

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