TY - JOUR
T1 - Behavior of a cross-linked attachment site
T2 - Testing the role of branch migration in site-specific recombination
AU - Cowart, Marlon
AU - Benkovic, Stephen J.
AU - Nash, Howard A.
N1 - Funding Information:
Andrew Granston, Richard Gumport and Robert Weisberg provided valuable comments on this manuscript. We thank Howard Benjamin for advice on labeling protocols, Todd Capson for T4 DNA polymerase, Cathy Joyce for the plasmid overproducer of the Y66S era-mutant of Klenow fragment and Carol Robertson for purified recombination proteins. We are also grateful to Marshal Nirenberg for use of the Betagen instrument and to Monese Christensen for preparation of this manuscript. This work was funded in part by a grant to S.J.B. from the National Institute of General Medical Sciences; M.C. was supported in part by a National Research Service Award from the National Institute of General Medical Sciences.
PY - 1991/8/5
Y1 - 1991/8/5
N2 - Integrative recombination of bacteriophage lambda requires perfect homology between partners over a short segment of DNA, the overlap region, that separates the positions of top and bottom strand exchange. We constructed a specific cross-link between complementary strands in the overlap region of one partner, using a method designed to introduce minimal distortion of DNA. The modified attachment site could initiate recombination, forming a Holliday junction, but could not resolve this junction so as to complete the recombination. This demonstrates that the ability of complementary base-pairs to dissociate is important for overlap region function and strongly supports the view that branch migration across this region is the way homology is sensed during integrative recombination.
AB - Integrative recombination of bacteriophage lambda requires perfect homology between partners over a short segment of DNA, the overlap region, that separates the positions of top and bottom strand exchange. We constructed a specific cross-link between complementary strands in the overlap region of one partner, using a method designed to introduce minimal distortion of DNA. The modified attachment site could initiate recombination, forming a Holliday junction, but could not resolve this junction so as to complete the recombination. This demonstrates that the ability of complementary base-pairs to dissociate is important for overlap region function and strongly supports the view that branch migration across this region is the way homology is sensed during integrative recombination.
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U2 - 10.1016/0022-2836(91)90105-F
DO - 10.1016/0022-2836(91)90105-F
M3 - Article
C2 - 1831237
AN - SCOPUS:0025894959
VL - 220
SP - 621
EP - 629
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
SN - 0022-2836
IS - 3
ER -