Topological characterization of the DnaA-oriC complex using single-molecule nanomanipuation.

Nucleic Acids Research
Sylvain ZormanT Strick

Abstract

In most bacteria, the timing and synchrony of initiation of chromosomal replication are determined by the binding of the AAA(+) protein DnaA to a set of high- and low-affinity sites found within the origin of chromosomal replication (oriC). Despite the large amount of information on the role and regulation of DnaA, the actual structure of the DnaA-oriC complex and the mechanism by which it primes the origin for the initiation of replication remain unclear. In this study, we have performed magnetic tweezers experiments to investigate the structural properties of the DnaA-oriC complex. We show that the DnaA-ATP-oriC complex adopts a right-handed helical conformation involving a variable amount of DNA and protein whose features fit qualitatively as well as quantitatively with an existing model based on the crystal structure of a truncated DnaA tetramer obtained in the absence of DNA. We also investigate the topological effect of oriC's DNA unwinding element.

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Citations

Jun 12, 2013·Annual Review of Biochemistry·Alessandro CostaJames M Berger
Apr 22, 2014·Journal of Molecular Biology·Rafał DonczewAnna Zawilak-Pawlik
Feb 4, 2014·Biophysical Chemistry·E A GalburtA Ruiz Manzano
Jun 18, 2015·Frontiers in Microbiology·Alan C Leonard, Julia E Grimwade
Jan 23, 2015·Frontiers in Microbiology·Marcin WolańskiJolanta Zakrzewska-Czerwińska
Dec 6, 2014·Nucleic Acids Research·Hsin-Mei ChengMichael Schlierf
Oct 18, 2016·Frontiers in Molecular Biosciences·Jyoti K JhaDhruba K Chattoraj
Jan 12, 2017·Genes·Katie H Jameson, Anthony J Wilkinson
Apr 4, 2013·Nature Reviews. Microbiology·Andrew Robinson, Antoine M van Oijen
Sep 13, 2019·PLoS Genetics·Babatunde Ekundayo, Franziska Bleichert
Jul 4, 2019·The EMBO Journal·Tomas T RichardsonHeath Murray
Jan 14, 2020·Nucleic Acids Research·Carolina CarrascoFernando Moreno-Herrero
May 26, 2018·Nucleic Acids Research·Julia E GrimwadeAlan C Leonard
Sep 27, 2019·Frontiers in Microbiology·Alan C LeonardJulia E Grimwade
Nov 3, 2020·Frontiers in Microbiology·Ana M Oliveira PaivaWiep Klaas Smits

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Methods Mentioned

BETA
electron microscopy
magnetic tweezers
gel filtration

Software Mentioned

RIDA

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