The structure of supercoiled intermediates in DNA replication

Brian J. Peter, Chris Ullsperger, Hiroshi Hiasa, Kenneth J. Marians, Nicholas R. Cozzarelli

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113 Scopus citations

Abstract

We studied the structure of replication intermediates accumulated by Tus-induced arrest of plasmid DNA replication at termination sites. For intermediates generated both in vitro with purified components and in vivo, superhelical stress is distributed throughout the entire partially replicated molecule; daughter DNA segments are wound around each other, and the unreplicated region is supercoiled. Thus, unlinking of parental DNA strands by topoisomerases can be carried out both behind and in front of the replication fork. We explain why previous studies with prokaryotic and eukaryotic replication intermediates discerned only supercoiling in the unreplicated portion.

Original languageEnglish (US)
Pages (from-to)819-827
Number of pages9
JournalCell
Volume94
Issue number6
DOIs
StatePublished - Sep 18 1998

Bibliographical note

Funding Information:
We thank Alex Vologodskii for his valuable advice and Tom Hill for promptly sharing plasmids. B. J. P. was supported in part by a NIEHS training grant, and C. J. U. was supported by a Howard Hughes Medical Institute predoctoral fellowship. This work was supported by NIH grants to N. R. C. and K. J. M.

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