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DNA sequence determinant for FIp-induced DNA bending

K H Luetke¹, P D Sadowski

¹Department of Medical Genetics and Microbiology, University of Toronto, Ontario, Canada.

Molecular Microbiology

|August 14, 1998

Summary

The central AT basepair in the Flp recognition target (FRT) site is crucial for DNA bending by Flp recombinase. Altering this basepair changes DNA bend position and impacts recombination efficiency.

Area of Science:

Molecular Biology
Biochemistry
Genetics

Background:

Flp recombinase from Saccharomyces cerevisiae mediates site-specific recombination.
The Flp recognition target (FRT) site contains essential DNA sequences for Flp binding and activity.
DNA bending is a key feature of Flp-DNA interactions.

Purpose of the Study:

To investigate the role of the central AT basepair in the FRT site.
To determine how modifications to the core region affect Flp-induced DNA bending.
To understand the impact of altered DNA structure on Flp-mediated recombination.

Main Methods:

Site-directed mutagenesis of the FRT core region.
Analysis of DNA bending in Flp-DNA complexes.
Assays for Flp-mediated DNA cleavage and recombination proficiency.

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Main Results:

Replacing the central AT basepair with CG shifted the DNA bend location.
The central AT basepair acts as a flexure point for Flp-induced DNA bending.
Mutations in the core region altered strand cleavage and reduced overall recombination efficiency.

Conclusions:

The central AT basepair is critical for positioning DNA bends by dimeric Flp.
The DNA sequence and structure of the FRT core directly influence Flp recombinase function.
Understanding FRT site structure provides insights into site-specific recombination mechanisms.