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Functional requirements for heat induced genome amplification in Escherichia coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
35472 45091 2008 9 PDF Available
Title
Functional requirements for heat induced genome amplification in Escherichia coli
Abstract

A temperature shift-up induces extra rounds of fully replicated chromosomes in Escherichia coli and leads to an increase in DNA/mass ratio. In the present work we characterize the requirements for this heat-induced replication (HIR) with respect to replisome components, replication restart, and recombination functions. We found that HIR requires Klenow and 5′–3′ exonuclease activities from Pol I and Pol III, but does not require translesion synthesis polymerases. We also found that replication restart is PriA–PriB pathway dependent. The dnaC809 allele suppresses the dependency on PriA, confirming the requirement for primosome assembly, in which PriA helicase function is not required. Rep helicase and the replication-associated function of RecA were found to be essential. HIR has low recombination requirements, and no DSBs are generated by heat stress. We propose that the Pol I-dependent replisome in HIR, which gives a slow replication speed, is more unstable and disassembles more frequently than normal replisome. Rep and RecA would be required to stabilize HIR replication forks. Since neither D-loops nor R-loops support HIR restart, the PriA–PriB pathway must reload the replisome by a direct restart mechanism from a 3′-end of a nascent leading strand.

Keywords
Replication; Heat stress; Escherichia coli; Pol I; PriA; RecA
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Functional requirements for heat induced genome amplification in Escherichia coli
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 43, Issue 10, October 2008, Pages 1162–1170
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us