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M701559200v1
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Papers In Press, published online ahead of print April 23, 2007
J. Biol. Chem, 10.1074/jbc.M701559200
Submitted on February 21, 2007
Revised on April 23, 2007
Accepted on April 23, 2007

Replication fork reversal occurs spontaneously after digestion but is constrained in supercoiled domains

Marta Fierro-Fernández, Pablo Hernández, Dora B. Krimer, and Jorge B. Schvartzman

Biologia Celular y del Desarrollo, Centro de Investigaciones Biologicas (CSIC), Madrid 28040

Corresponding Author: schvartzman{at}cib.csic.es

Replication fork reversal (RFR) was investigated in undigested and linearized replication intermediates (RIs) of bacterial DNA plasmids containing a stalled fork. Two-dimensional (2D) agarose gel electrophoresis, a branch migration and extrusion assay, electron microscopy (EM) and DNA-psoralen crosslinking were used to show that extensive RFR and extrusion of the nascent-nascent duplex occurs spontaneously after DNA nicking and restriction enzyme digestion but fork retreat is severely limited in covalently closed supercoiled domains.


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[Abstract] [Full Text] [PDF]


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