Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (845K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (22)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Olavarrieta, L.
Right arrow Articles by Schvartzman, J. B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Olavarrieta, L.
Right arrow Articles by Schvartzman, J. B.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2002, Vol. 30, No. 3 656-666
© 2002 Oxford University Press

Supercoiling, knotting and replication fork reversal in partially replicated plasmids

L. Olavarrieta, M. L. Martínez-Robles, J. M. Sogo1, A. Stasiak2, P. Hernández, D. B. Krimer and J. B. Schvartzman*

Departamento de Biología Celular y del Desarrollo, Centro de Investigaciones Biológicas (CSIC), Velázquez 144, 28006 Madrid, Spain, 1Institut für Zellbiologie ETH-Hönggerberg, CH-8093 Zürich, Switzerland and 2Laboratoire d’Analyse Ultrastructurale, Bâtiment de Biologie, Université de Lausanne, CH-1015 Lausanne, Switzerland

To study the structure of partially replicated plasmids, we cloned the Escherichia coli polar replication terminator TerE in its active orientation at different locations in the ColE1 vector pBR18. The resulting plasmids, pBR18-TerE@StyI and pBR18-TerE@EcoRI, were analyzed by neutral/neutral two-dimensional agarose gel electrophoresis and electron microscopy. Replication forks stop at the Ter–TUS complex, leading to the accumulation of specific replication intermediates with a mass 1.26 times the mass of non-replicating plasmids for pBR18-TerE@StyI and 1.57 times for pBR18-TerE@EcoRI. The number of knotted bubbles detected after digestion with ScaI and the number and electrophoretic mobility of undigested partially replicated topoisomers reflect the changes in plasmid topology that occur in DNA molecules replicated to different extents. Exposure to increasing concentrations of chloroquine or ethidium bromide revealed that partially replicated topoisomers (CCCRIs) do not sustain positive supercoiling as efficiently as their non-replicating counterparts. It was suggested that this occurs because in partially replicated plasmids a positive {Delta}Lk is absorbed by regression of the replication fork. Indeed, we showed by electron microscopy that, at least in the presence of chloroquine, some of the CCCRIs of pBR18-Ter@StyI formed Holliday-like junction structures characteristic of reversed forks. However, not all the positive supercoiling was absorbed by fork reversal in the presence of high concentrations of ethidium bromide.

* To whom correspondence should be addressed. Tel: +34 91 5644562; Fax: +34 91 564 8749; Email: schvartzman{at}cib.csic.es


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
M. L. Martinez-Robles, G. Witz, P. Hernandez, J. B. Schvartzman, A. Stasiak, and D. B. Krimer
Interplay of DNA supercoiling and catenation during the segregation of sister duplexes
Nucleic Acids Res., August 1, 2009; 37(15): 5126 - 5137.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
J. Atkinson and P. McGlynn
Replication fork reversal and the maintenance of genome stability
Nucleic Acids Res., June 1, 2009; 37(11): 3475 - 3492.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Y. Burnier, J. Dorier, and A. Stasiak
DNA supercoiling inhibits DNA knotting
Nucleic Acids Res., September 1, 2008; 36(15): 4956 - 4963.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
I. K. Darcy, R. G. Scharein, and A. Stasiak
3D visualization software to analyze topological outcomes of topoisomerase reactions
Nucleic Acids Res., June 1, 2008; 36(11): 3515 - 3521.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Y. Burnier, C. Weber, A. Flammini, and A. Stasiak
Local selection rules that can determine specific pathways of DNA unknotting by type II DNA topoisomerases
Nucleic Acids Res., August 1, 2007; (2007) gkm532v1.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
E. Guarino, I. Salguero, A. Jimenez-Sanchez, and E. C. Guzman
Double-Strand Break Generation under Deoxyribonucleotide Starvation in Escherichia coli
J. Bacteriol., August 1, 2007; 189(15): 5782 - 5786.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Fierro-Fernandez, P. Hernandez, D. B. Krimer, and J. B. Schvartzman
Replication Fork Reversal Occurs Spontaneously after Digestion but Is Constrained in Supercoiled Domains
J. Biol. Chem., June 22, 2007; 282(25): 18190 - 18196.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
M. Fierro-Fernandez, P. Hernandez, D. B. Krimer, A. Stasiak, and J. B. Schvartzman
From the Cover: Topological locking restrains replication fork reversal
PNAS, January 30, 2007; 104(5): 1500 - 1505.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
B. Carrasco, M. C. Cozar, R. Lurz, J. C. Alonso, and S. Ayora
Genetic Recombination in Bacillus subtilis 168: Contribution of Holliday Junction Processing Functions in Chromosome Segregation
J. Bacteriol., September 1, 2004; 186(17): 5557 - 5566.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
B. Michel, G. Grompone, M.-J. Flores, and V. Bidnenko
Multiple pathways process stalled replication forks
PNAS, August 31, 2004; 101(35): 12783 - 12788.
[Abstract] [Full Text] [PDF]

Home page
 GeneticsHome page
J. R. Donaldson, C. T. Courcelle, and J. Courcelle
RuvAB and RecG Are Not Essential for the Recovery of DNA Synthesis Following UV-Induced DNA Damage in Escherichia coli
Genetics, April 1, 2004; 166(4): 1631 - 1640.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
A. Benguria, P. Hernandez, D. B. Krimer, and J. B. Schvartzman
Sir2p suppresses recombination of replication forks stalled at the replication fork barrier of ribosomal DNA in Saccharomyces cerevisiae
Nucleic Acids Res., February 1, 2003; 31(3): 893 - 898.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.