DNA-replication recovery inhibition and subsequent reinitiation in UV-radiation-damaged E. coli: a strategy for survival

Electron microscopy of replicating intermediates has been quite useful in understanding the mechanism of DNA replication in DNA molecules of bacteriophage, mitochondria and plasmids. The use of partial denaturation mapping has made the tool more powerful by providing a frame of reference by which the position of the replicating forks in bacteriophage DNA can be determined on the circular replicating molecules. This provided an easy means to find the origin and direction of replication in λ and P2 phage DNA molecules. DNA of temperate E. coli phage 186 was found to have an unique denaturation map and encouraged us to look into its mode of replication.

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E. coli primase and DNA polymerase III holoenzyme are able to bind concurrently to a primed template during DNA replication

Scientific Reports ◽

10.1038/s41598-019-51031-0 ◽

2019 ◽

Vol 9 (1) ◽

Author(s):

Andrea Bogutzki ◽

Natalie Naue ◽

Lidia Litz ◽

Andreas Pich ◽

Ute Curth

Keyword(s):

Dna Replication ◽

Dna Polymerase ◽

Protein Interactions ◽

Dna Polymerase Iii ◽

Protein Protein Interactions ◽

Single Stranded Dna ◽

E Coli ◽

Polymerase Iii ◽

C Terminus ◽

Pol Iii

Abstract During DNA replication in E. coli, a switch between DnaG primase and DNA polymerase III holoenzyme (pol III) activities has to occur every time when the synthesis of a new Okazaki fragment starts. As both primase and the χ subunit of pol III interact with the highly conserved C-terminus of single-stranded DNA-binding protein (SSB), it had been proposed that the binding of both proteins to SSB is mutually exclusive. Using a replication system containing the origin of replication of the single-stranded DNA phage G4 (G4ori) saturated with SSB, we tested whether DnaG and pol III can bind concurrently to the primed template. We found that the addition of pol III does not lead to a displacement of primase, but to the formation of higher complexes. Even pol III-mediated primer elongation by one or several DNA nucleotides does not result in the dissociation of DnaG. About 10 nucleotides have to be added in order to displace one of the two primase molecules bound to SSB-saturated G4ori. The concurrent binding of primase and pol III is highly plausible, since even the SSB tetramer situated directly next to the 3′-terminus of the primer provides four C-termini for protein-protein interactions.

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The Large Subunit of Replication Factor C (Rfclp/Cdc44p) Is Required for DNA Replication and DNA Repair in Saccharomyces cerevisiae

Genetics ◽

10.1093/genetics/142.1.65 ◽

1996 ◽

Vol 142 (1) ◽

pp. 65-78 ◽

Cited By ~ 7

Author(s):

Michael A McAlear ◽

K Michelle Tuffo ◽

Connie Holm

Keyword(s):

Saccharomyces Cerevisiae ◽

Dna Repair ◽

Dna Replication ◽

Uv Radiation ◽

Large Subunit ◽

Restrictive Temperature ◽

Replication Factor C ◽

Replication Factor ◽

Factor C

We used genetic and biochemical techniques to characterize the phenotypes associated with mutations affecting the large subunit of replication factor C (Cdc44p or Rfc1p) in Saccharomyces cerevisiae. We demonstrate that Cdc44p is required for both DNA replication and DNA repair in vivo. Cold-sensitive cdc44 mutants experience a delay in traversing S phase at the restrictive temperature following alpha factor arrest; although mutant cells eventually accumulate with a G2/M DNA content, they undergo a cell cycle arrest and initiate neither mitosis nor a new round of DNA synthesis. cdc44 mutants also exhibit an elevated level of spontaneous mutation, and they are sensitive both to the DNA damaging agent methylmethane sulfonate and to exposure to UV radiation. After exposure to UV radiation, cdc44 mutants at the restrictive temperature contain higher levels of single-stranded DNA breaks than do wild-type cells. This observation is consistent with the hypothesis that Cdc44p is involved in repairing gaps in the DNA after the excision of damaged bases. Thus, Cdc44p plays an important role in both DNA replication and DNA repair in vivo.

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UV-induced Hyperphosphorylation of Replication Protein A Depends on DNA Replication and Expression of ATM Protein

Molecular Biology of the Cell ◽

10.1091/mbc.12.5.1199 ◽

2001 ◽

Vol 12 (5) ◽

pp. 1199-1213 ◽

Cited By ~ 58

Author(s):

Gregory G. Oakley ◽

Lisa I. Loberg ◽

Jiaqin Yao ◽

Mary A. Risinger ◽

Remy L. Yunker ◽

...

Keyword(s):

Dna Replication ◽

Uv Radiation ◽

Excision Repair ◽

Genetic Disorder ◽

Protein A ◽

Dna Recombination ◽

Delayed Response ◽

Replication Intermediates

Exposure to DNA-damaging agents triggers signal transduction pathways that are thought to play a role in maintenance of genomic stability. A key protein in the cellular processes of nucleotide excision repair, DNA recombination, and DNA double-strand break repair is the single-stranded DNA binding protein, RPA. We showed previously that the p34 subunit of RPA becomes hyperphosphorylated as a delayed response (4–8 h) to UV radiation (10–30 J/m2). Here we show that UV-induced RPA-p34 hyperphosphorylation depends on expression of ATM, the product of the gene mutated in the human genetic disorder ataxia telangiectasia (A-T). UV-induced RPA-p34 hyperphosphorylation was not observed in A-T cells, but this response was restored by ATM expression. Furthermore, purified ATM kinase phosphorylates the p34 subunit of RPA complex in vitro at many of the same sites that are phosphorylated in vivo after UV radiation. Induction of this DNA damage response was also dependent on DNA replication; inhibition of DNA replication by aphidicolin prevented induction of RPA-p34 hyperphosphorylation by UV radiation. We postulate that this pathway is triggered by the accumulation of aberrant DNA replication intermediates, resulting from DNA replication fork blockage by UV photoproducts. Further, we suggest that RPA-p34 is hyperphosphorylated as a participant in the recombinational postreplication repair of these replication products. Successful resolution of these replication intermediates reduces the accumulation of chromosomal aberrations that would otherwise occur as a consequence of UV radiation.

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Pseudovirulent mutants of λb221poriCasnA resulting from mutations in or near oriC, the E. coli origin of DNA replication

MGG Molecular & General Genetics ◽

10.1007/bf00270489 ◽

1980 ◽

Vol 178 (2) ◽

pp. 391-396 ◽

Cited By ~ 1

Author(s):

Larry Soll

Keyword(s):

Dna Replication ◽

E Coli ◽

Origin Of Dna Replication

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Dissecting the Control Mechanisms for DNA Replication and Cell Division in E. coli

Cell Reports ◽

10.1016/j.celrep.2018.09.061 ◽

2018 ◽

Vol 25 (3) ◽

pp. 761-771.e4 ◽

Cited By ~ 21

Author(s):

Gabriele Micali ◽

Jacopo Grilli ◽

Jacopo Marchi ◽

Matteo Osella ◽

Marco Cosentino Lagomarsino

Keyword(s):

Cell Division ◽

Dna Replication ◽

Control Mechanisms ◽

E Coli

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UV- and MMS-induced mutagenesis of lambdaO(am)8 phage under nonpermissive conditions for phage DNA replication.

Acta Biochimica Polonica ◽

10.18388/abp.2003_3624 ◽

2003 ◽

Vol 50 (4) ◽

pp. 921-939 ◽

Cited By ~ 1

Author(s):

Joanna Krwawicz ◽

Anna Czajkowska ◽

Magdalena Felczak ◽

Irena Pietrzykowska

Keyword(s):

Dna Replication ◽

Dna Polymerase ◽

Excision Repair ◽

Protein S ◽

Dna Lesions ◽

E Coli ◽

Phage Dna ◽

Induced Mutagenesis ◽

C Proteins ◽

Inducible Protein

Mutagenesis in Escherichia coli, a subject of many years of study is considered to be related to DNA replication. DNA lesions nonrepaired by the error-free nucleotide excision repair (NER), base excision repair (BER) and recombination repair (RR), stop replication at the fork. Reinitiation needs translesion synthesis (TLS) by DNA polymerase V (UmuC), which in the presence of accessory proteins, UmuD', RecA and ssDNA-binding protein (SSB), has an ability to bypass the lesion with high mutagenicity. This enables reinitiation and extension of DNA replication by DNA polymerase III (Pol III). We studied UV- and MMS-induced mutagenesis of lambdaO(am)8 phage in E. coli 594 sup+ host, unable to replicate the phage DNA, as a possible model for mutagenesis induced in nondividing cells (e.g. somatic cells). We show that in E. coli 594 sup+ cells UV- and MMS-induced mutagenesis of lambdaO(am)8 phage may occur. This mutagenic process requires both the UmuD' and C proteins, albeit a high level of UmuD' and low level of UmuC seem to be necessary and sufficient. We compared UV-induced mutagenesis of lambdaO(am)8 in nonpermissive (594 sup+) and permissive (C600 supE) conditions for phage DNA replication. It appeared that while the mutagenesis of lambdaO(am)8 in 594 sup+ requires the UmuD' and C proteins, which can not be replaced by other SOS-inducible protein(s), in C600 supE their functions may be replaced by other inducible protein(s), possibly DNA polymerase IV (DinB). Mutations induced under nonpermissive conditions for phage DNA replication are resistant to mismatch repair (MMR), while among those induced under permissive conditions, only about 40% are resistant.

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A Molecular Mousetrap Determines Polarity of Termination of DNA Replication in E. coli

Cell ◽

10.1016/j.cell.2006.04.040 ◽

2006 ◽

Vol 125 (7) ◽

pp. 1309-1319 ◽

Cited By ~ 83

Author(s):

Mark D. Mulcair ◽

Patrick M. Schaeffer ◽

Aaron J. Oakley ◽

Hannah F. Cross ◽

Cameron Neylon ◽

...

Keyword(s):

Dna Replication ◽

E Coli

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Solar UV damage to cellular DNA: from mechanisms to biological effects

Photochemical & Photobiological Sciences ◽

10.1039/c8pp00182k ◽

2018 ◽

Vol 17 (12) ◽

pp. 1842-1852 ◽

Cited By ~ 25

Author(s):

Leon H. F. Mullenders

Keyword(s):

Dna Replication ◽

Uv Radiation ◽

Biological Effects ◽

Uv Damage ◽

Solar Uv ◽

Cellular Dna ◽

Solar Ultraviolet

Solar ultraviolet (UV) radiation generates bulky photodimers at di-pyrimidine sites that pose stress to cells and organisms by hindering DNA replication and transcription.

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