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Stimulation of IS1 excision by bacteriophage P1 ref function.

S D Lu¹, D Lu, M Gottesman

¹Laboratory of Molecular Biology, National Cancer Institute, Bethesda, Maryland 20205.

Journal of Bacteriology

|June 1, 1989

Summary

Coliphage P1 lysogenization enhances galT::IS1 reversion via microhomologous recombination. The P1 ref gene product stimulates precise DNA excision, dependent on RecA activity.

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Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

Coliphage P1 lysogenization influences host DNA.
IS1 insertion mutations can revert through recombination.

Purpose of the Study:

Investigate the mechanism of galT::IS1 reversion during P1 lysogenization.
Identify and characterize the P1 gene responsible for this phenomenon.

Main Methods:

Cloning and sequencing of the P1 ref gene.
Construction of a ref mutant.
Assessing Ref protein activity in various bacterial mutants (recB, himA, recA, lexA3).

Main Results:

P1 lysogenization with P1c1.100 significantly increased galT::IS1 reversion frequency.
The P1 ref gene was identified, encoding a 22.8 kDa protein.
Ref protein stimulates precise excision of IS1 in recB and himA mutants, but not recA mutants.
Ref activity is independent of lexA3, suggesting direct RecA involvement.
A ref::Tn10 mutant showed no significant defect in lytic growth or lysogenization.

Conclusions:

The P1 ref gene product mediates microhomologous recombination, facilitating IS1 excision.
Ref's role in P1 physiology requires further investigation.
The findings highlight phage-host interactions in DNA repair and recombination.

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