DnaA box sequences as the site for helicase delivery during plasmid RK2 replication initiation in Escherichia coli

The Journal of Biological Chemistry
Marcin PacekIgor Konieczny

Abstract

DnaA box sequences are a common motif present within the replication origin region of a diverse group of bacteria and prokaryotic extrachromosomal genetic elements. Although the origin opening caused by binding of the host DnaA protein has been shown to be critical for the loading of the DnaB helicase, to date there has been no direct evidence presented for the formation of the DnaB complex at the DnaA box site. For these studies, we used the replication origin of plasmid RK2 (oriV), containing a cluster of four DnaA boxes that bind DnaA proteins isolated from different bacterial species (Caspi, R., Helinski, D. R., Pacek, M., and Konieczny, I. (2000) J. Biol. Chem. 275, 18454-18461). Size exclusion chromatography, surface plasmon resonance, and electron microscopy experiments demonstrated that the DnaB helicase is delivered to the DnaA box region, which is localized approximately 200 base pairs upstream from the region of origin opening and a potential site for helicase entry. The DnaABC complex was formed on both double-stranded superhelical and linear RK2 templates. A strict DnaA box sequence requirement for stable formation of that nucleoprotein structure was confirmed. In addition, our experiments provide evidence for inte...Continue Reading

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Citations

Dec 26, 2002·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Jan 14, 2003·EMBO Reports·Igor Konieczny
Nov 8, 2002·Molecular Microbiology·Christoph Weigel, Harald Seitz
Aug 8, 2008·Proceedings of the National Academy of Sciences of the United States of America·Magdalena RajewskaIgor Konieczny
Jan 5, 2006·Applied and Environmental Microbiology·Ju-Hoon Lee, Daniel J O'Sullivan
Jul 2, 2003·Proceedings of the National Academy of Sciences of the United States of America·Yong JiangAresa Toukdarian
Dec 3, 2014·Plasmid·Elzbieta ZabrockaIgor Konieczny
Jul 22, 2015·Proceedings of the National Academy of Sciences of the United States of America·Aleksandra WawrzyckaIgor Konieczny
Jul 6, 2010·Plasmid·Katarzyna KolatkaIgor Konieczny
Dec 17, 2015·Current Microbiology·Lilian C CarneiroLuiz Artur M Bataus
Nov 19, 2011·FEMS Microbiology Reviews·Magdalena RajewskaIgor Konieczny
Aug 27, 2016·Frontiers in Molecular Biosciences·Katarzyna E WegrzynIgor Konieczny
Mar 15, 2003·The Journal of Biological Chemistry·Teresa Díaz-LópezRafael Giraldo
Jun 25, 2015·Microbiology Spectrum·Igor KoniecznyKatarzyna Wegrzyn

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