Mutational Analysis Reveals Escherichia Coli OriC Interacts with Both DnaA-ATP and DnaA-ADP During Pre-RC Assembly

Overview

Journal Mol Microbiol

Specialties Microbiology
Molecular Biology

Date 2007 Sep 14

PMID 17850252

Citations 24

Authors

Julia E Grimwade

Julien J-C Torgue

Kevin C McGarry

Tanya Rozgaja

Sareena T Enloe

Alan C Leonard

Affiliations

Soon will be listed here.

Abstract

Prior to initiating DNA synthesis, Escherichia coli oriC switches from ORC, comprising initiator DnaA bound at three high-affinity sites, to pre-RC, when additional DnaA molecules interact with low-affinity sites. Two types of low-affinity sites exist: R boxes that bind DnaA-ATP and DnaA-ADP with equal affinity, and I-sites with a three- to fourfold preference for DnaA-ATP. To assess the regulatory role of weak DnaA interactions during pre-RC assembly in vivo, we compared the behaviour of plasmid-borne wild-type oriC with mutants having an increased or decreased number of DnaA-ATP discriminatory I-sites. Increasing the number of discriminatory sites by replacing R5M with I2 inactivated extrachromosomal oriC function. Mutants with no discriminatory sites perturbed host growth and rapidly replaced wild-type chromosomal oriC, but normal function returned if one I-site was restored at either the I2, I3 or R5M position. These observations are consistent with assembly of E. coli pre-RC in vivo from mixtures of DnaA-ATP and DnaA-ADP, with I-site interactions coupling pre-RC assembly to DnaA-ATP levels.

Citing Articles

Blocking, Bending, and Binding: Regulation of Initiation of Chromosome Replication During the Cell Cycle by Transcriptional Modulators That Interact With Origin DNA.

Grimwade J, Leonard A Front Microbiol. 2021; 12:732270.

PMID: 34616385 PMC: 8488378. DOI: 10.3389/fmicb.2021.732270.

Putative Cooperative ATP-DnaA Binding to Double-Stranded DnaA Box and Single-Stranded DnaA-Trio Motif upon Replication Initiation Complex Assembly.

Jaworski P, Zyla-Uklejewicz D, Nowaczyk-Cieszewska M, Donczew R, Mielke T, Weigel C Int J Mol Sci. 2021; 22(12).

PMID: 34205762 PMC: 8235120. DOI: 10.3390/ijms22126643.

A nucleotide-dependent oligomerization of the Escherichia coli replication initiator DnaA requires residue His136 for remodeling of the chromosomal origin.

Saxena R, Stanley C, Kumar P, Cuneo M, Patil D, Jha J Nucleic Acids Res. 2019; 48(1):200-211.

PMID: 31665475 PMC: 7145717. DOI: 10.1093/nar/gkz939.

Changing Perspectives on the Role of DnaA-ATP in Orisome Function and Timing Regulation.

Leonard A, Rao P, Kadam R, Grimwade J Front Microbiol. 2019; 10:2009.

PMID: 31555240 PMC: 6727663. DOI: 10.3389/fmicb.2019.02009.

Blocking the Trigger: Inhibition of the Initiation of Bacterial Chromosome Replication as an Antimicrobial Strategy.

Grimwade J, Leonard A Antibiotics (Basel). 2019; 8(3).

PMID: 31390740 PMC: 6784150. DOI: 10.3390/antibiotics8030111.

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