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J. Biol. Chem., Vol. 275, Issue 44, 34757-34765, November 3, 2000
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From the Molecular Biology Program, Memorial Sloan-Kettering Cancer
Center, New York, New York 10021
There are two modes of DNA synthesis at a
replication fork. The leading strand is synthesized in a continuous
fashion in lengths that in Escherichia coli can be in
excess of 2 megabases. On the other hand, the lagging strand is
synthesized in relatively short stretches of 2 kilobases.
Nevertheless, identical assemblies of the DNA polymerase III core
tethered to the
sliding clamp account for both modes of DNA
synthesis. Yet the same lagging strand polymerase accounts for the
synthesis of all Okazaki fragments at a replication fork, cycling
repeatedly every 1 or 2 s from the 3'-end of the just-completed
fragment to the 3'-end of the new primer. Several models have been
invoked to account for the rapid cycling of a polymerase complex that
can remain bound to the template for upward of 40 min. By using
isolated replication protein-DNA template complexes, we have tested
these models and show here that cycling of the lagging strand
polymerase can be triggered by either the action of primase binding to
the replisome and synthesizing a primer or by collision of the lagging
strand polymerase with the 5'-end of the previous Okazaki fragment.
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