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J. Biol. Chem., Vol. 277, Issue 16, 14321-14328, April 19, 2002
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and§¶
From the Recombination-dependent replication
is an essential housekeeping function in prokaryotes and eukaryotes,
serving, for example, to restart DNA replication after the repair of a
double-strand break. Little is known about the interplay between the
recombination and replication machinery when recombination
intermediates are used as substrates for DNA replication. We show here
that recombination intermediates formed between linear duplex and
supercoiled plasmid DNAs are substrates for a generalized strand
displacement DNA synthesis reaction in which the 3'-OH of the invading
strand in the recombination intermediate is used as a primer. DNA
synthesis is driven by negative superhelicity and is inhibited if
disassembly of the RecA filament is prevented. Thus, assembly and
disassembly of RecA filaments in the same direction facilitates
filament clearance from the 3'-end of the invading strand,
allowing DNA synthesis to occur from recombination intermediates.
Graduate Program in Biochemistry and
Structural Biology, Graduate School of Medical Sciences of the Weill
College of Medicine of Cornell University and § Molecular
Biology Program, Memorial Sloan-Kettering Cancer Center,
New York, New York 10021
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