Revised model for the DNA replication fork in Bacillus subtilis: polymerase asymmetry

Replication of DNA is a vital process for growth and cell division in all domains of life. The mechanisms of helicase loading as well as hand-off of nascent RNA primers from the primase to the replicative polymerases during DNA synthesis are of fundamental importance. The prototype structural/funct...

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Main Author: Paschalis, Vasileios
Format: Thesis (University of Nottingham only)
Language:English
Published: 2017
Online Access:https://eprints.nottingham.ac.uk/40887/
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author Paschalis, Vasileios
author_facet Paschalis, Vasileios
author_sort Paschalis, Vasileios
building Nottingham Research Data Repository
collection Online Access
description Replication of DNA is a vital process for growth and cell division in all domains of life. The mechanisms of helicase loading as well as hand-off of nascent RNA primers from the primase to the replicative polymerases during DNA synthesis are of fundamental importance. The prototype structural/functional model provided by the widely studied Gram-negative Escherichia coli DNA replication system is not universally applicable. The current model for the Gram-positive Bacillus subtilis DNA replication suggests that PolC is the main processive replicative polymerase with DnaE playing an essential but minor role during DNA synthesis. Our results suggest that DnaE is a major polymerase important for DNA replication and DNA repair. DnaE polymerase activity is stimulated in the presence of SSB, the clamp DnaN and PolC polymerase, its error-prone synthesis is modulated by the aforementioned proteins and its errors are corrected in trans by the PolC exonuclease domain, in a template specific ternary DNA-DnaE-PolC complex. Hereby, we propose a new revised model for DNA replication in Bacillus subtilis, where DnaE is the major replicative polymerase on the lagging strand and PolC, which is the major polymerase on the leading strand, provides the crucial proof-reading activity in trans. These findings revise the current model in Bacillus subtilis and suggest a division of labour between DnaE and PolC polymerases.
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format Thesis (University of Nottingham only)
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spelling nottingham-408872025-02-28T13:41:54Z https://eprints.nottingham.ac.uk/40887/ Revised model for the DNA replication fork in Bacillus subtilis: polymerase asymmetry Paschalis, Vasileios Replication of DNA is a vital process for growth and cell division in all domains of life. The mechanisms of helicase loading as well as hand-off of nascent RNA primers from the primase to the replicative polymerases during DNA synthesis are of fundamental importance. The prototype structural/functional model provided by the widely studied Gram-negative Escherichia coli DNA replication system is not universally applicable. The current model for the Gram-positive Bacillus subtilis DNA replication suggests that PolC is the main processive replicative polymerase with DnaE playing an essential but minor role during DNA synthesis. Our results suggest that DnaE is a major polymerase important for DNA replication and DNA repair. DnaE polymerase activity is stimulated in the presence of SSB, the clamp DnaN and PolC polymerase, its error-prone synthesis is modulated by the aforementioned proteins and its errors are corrected in trans by the PolC exonuclease domain, in a template specific ternary DNA-DnaE-PolC complex. Hereby, we propose a new revised model for DNA replication in Bacillus subtilis, where DnaE is the major replicative polymerase on the lagging strand and PolC, which is the major polymerase on the leading strand, provides the crucial proof-reading activity in trans. These findings revise the current model in Bacillus subtilis and suggest a division of labour between DnaE and PolC polymerases. 2017-07-18 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/40887/1/Revised%20Thesis-Vasileios%20Paschalis.pdf Paschalis, Vasileios (2017) Revised model for the DNA replication fork in Bacillus subtilis: polymerase asymmetry. PhD thesis, University of Nottingham.
spellingShingle Paschalis, Vasileios
Revised model for the DNA replication fork in Bacillus subtilis: polymerase asymmetry
title Revised model for the DNA replication fork in Bacillus subtilis: polymerase asymmetry
title_full Revised model for the DNA replication fork in Bacillus subtilis: polymerase asymmetry
title_fullStr Revised model for the DNA replication fork in Bacillus subtilis: polymerase asymmetry
title_full_unstemmed Revised model for the DNA replication fork in Bacillus subtilis: polymerase asymmetry
title_short Revised model for the DNA replication fork in Bacillus subtilis: polymerase asymmetry
title_sort revised model for the dna replication fork in bacillus subtilis: polymerase asymmetry
url https://eprints.nottingham.ac.uk/40887/