Nucleic Acids Research, Vol 26, Issue 6 1401-1407, Copyright © 1998 by Oxford University Press
KH Luetke and PD Sadowski
The Flp site-specific recombinase from Saccharomyces cerevisiae induces DNA
bending upon interaction with the Flp recognition target (FRT) site. The
minimal FRT site is comprised of two inverted binding elements which flank
a central core region. Binding of a single monomer of Flp to DNA induces a
DNA bend of 60 degrees. The position of this bend differed depending on
whether the substrate contained a single binding element or a two-element
FRT site. In the present work we tested and disproved a model in which a
single Flp monomer interacts with both symmetry elements of a single FRT
site. Likewise, we showed that a model in which a Flp monomer dissociates
from a singly occupied FRT site and reassociates with the unbound element
of another singly occupied FRT site during electrophoresis, does not
account for the apparent shift in the position of the bend centre. It seems
that the movement of a Flp monomer between the a and b elements of one FRT
site during electrophoresis accounts for this anomaly. The position of the
DNA bend resulting from the association of a Flp monomer with the FRT site
is also influenced by the DNA sequences flanking the site. We conclude that
attempts to measure the bend centre of a complex of one Flp molecule bound
to a DNA containing two binding elements give misleading results. The
position of the bend is more accurately measured in the presence of a
single binding element.
ARTICLES
Determinants of the position of a Flp-induced DNA bend
Department of Medical Genetics and Microbiology, Medical Sciences Building, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
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