scholarly journals REV1 restrains DNA polymerase ζ to ensure frame fidelity during translesion synthesis of UV photoproducts in vivo

2008 ◽  
Vol 36 (21) ◽  
pp. 6767-6780 ◽  
Author(s):  
Dávid Szüts ◽  
Adam P. Marcus ◽  
Masayuki Himoto ◽  
Shigenori Iwai ◽  
Julian E. Sale
Keyword(s):  
Dna Polymerase ◽  
Translesion Synthesis ◽  
Uv Photoproducts

scholarly journals Analysis of Damage Tolerance Pathways in Saccharomyces cerevisiae: a Requirement for Rev3 DNA Polymerase in Translesion Synthesis

1998 ◽  
Vol 18 (2) ◽  
pp. 960-966 ◽  
Author(s):  
K. Baynton ◽  
A. Bresson-Roy ◽  
R. P. P. Fuchs
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Polymerase ◽  
Excision Repair ◽  
Translesion Synthesis ◽  
Hybridization Technique ◽  
Nucleotide Excision ◽  
Yeast Transformants ◽  
Plasmid Design ◽  
First Time

ABSTRACT The replication of double-stranded plasmids containing a singleN-2-acetylaminofluorene (AAF) adduct located in a short, heteroduplex sequence was analyzed in Saccharomyces cerevisiae. The strains used were proficient or deficient for the activity of DNA polymerase ζ (REV3 andrev3Δ, respectively) in a mismatch and nucleotide excision repair-defective background (msh2Δ rad10Δ). The plasmid design enabled the determination of the frequency with which translesion synthesis (TLS) and mechanisms avoiding the adduct by using the undamaged, complementary strand (damage avoidance mechanisms) are invoked to complete replication. To this end, a hybridization technique was implemented to probe plasmid DNA isolated from individual yeast transformants by using short, 32P-end-labeled oligonucleotides specific to each strand of the heteroduplex. In both the REV3 and rev3Δ strains, the two strands of an unmodified heteroduplex plasmid were replicated in ∼80% of the transformants, with the remaining 20% having possibly undergone prereplicative MSH2-independent mismatch repair. However, in the presence of the AAF adduct, TLS occurred in only 8% of theREV3 transformants, among which 97% was mostly error free and only 3% resulted in a mutation. All TLS observed in theREV3 strain was abolished in the rev3Δ mutant, providing for the first time in vivo biochemical evidence of a requirement for the Rev3 protein in TLS.

scholarly journals The processivity factor β controls DNA polymerase IV traffic during spontaneous mutagenesis and translesion synthesis in vivo

EMBO Reports ◽  
2002 ◽  
Vol 3 (1) ◽  
pp. 45-49 ◽  
Author(s):  
Nathalie Lenne‐Samuel ◽  
Jérôme Wagner ◽  
Hélène Etienne ◽  
Robert P P Fuchs
Keyword(s):  
Dna Polymerase ◽  
Translesion Synthesis ◽  
Spontaneous Mutagenesis ◽  
Processivity Factor ◽  
Dna Polymerase Iv

scholarly journals PrimPol is required for replication reinitiation after mtDNA damage

2017 ◽  
Vol 114 (43) ◽  
pp. 11398-11403 ◽  
Author(s):  
Rubén Torregrosa-Muñumer ◽  
Josefin M. E. Forslund ◽  
Steffi Goffart ◽  
Annika Pfeiffer ◽  
Gorazd Stojkovič ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Translesion Synthesis ◽  
Replication Forks ◽  
Mtdna Damage ◽  
Replicative Stress ◽  
Mtdna Replication ◽  
Stalled Replication Forks ◽  
Lagging Strand

Eukaryotic PrimPol is a recently discovered DNA-dependent DNA primase and translesion synthesis DNA polymerase found in the nucleus and mitochondria. Although PrimPol has been shown to be required for repriming of stalled replication forks in the nucleus, its role in mitochondria has remained unresolved. Here we demonstrate in vivo and in vitro that PrimPol can reinitiate stalled mtDNA replication and can prime mtDNA replication from nonconventional origins. Our results not only help in the understanding of how mitochondria cope with replicative stress but can also explain some controversial features of the lagging-strand replication.

scholarly journals DNA polymerase from temperate phage Bam35 is endowed with processive polymerization and abasic sites translesion synthesis capacity

2015 ◽  
Vol 112 (27) ◽  
pp. E3476-E3484 ◽  
Author(s):  
Mónica Berjón-Otero ◽  
Laurentino Villar ◽  
Miguel de Vega ◽  
Margarita Salas ◽  
Modesto Redrejo-Rodríguez
Keyword(s):  
Dna Polymerase ◽  
Biochemical Characterization ◽  
Multiple Displacement Amplification ◽  
Translesion Synthesis ◽  
Mobile Element ◽  
Genome Replication ◽  
Abasic Site ◽  
Abasic Sites

DNA polymerases (DNAPs) responsible for genome replication are highly faithful enzymes that nonetheless cannot deal with damaged DNA. In contrast, translesion synthesis (TLS) DNAPs are suitable for replicating modified template bases, although resulting in very low-fidelity products. Here we report the biochemical characterization of the temperate bacteriophage Bam35 DNA polymerase (B35DNAP), which belongs to the protein-primed subgroup of family B DNAPs, along with phage Φ29 and other viral and mobile element polymerases. B35DNAP is a highly faithful DNAP that can couple strand displacement to processive DNA synthesis. These properties allow it to perform multiple displacement amplification of plasmid DNA with a very low error rate. Despite its fidelity and proofreading activity, B35DNAP was able to successfully perform abasic site TLS without template realignment and inserting preferably an A opposite the abasic site (A rule). Moreover, deletion of the TPR2 subdomain, required for processivity, impaired primer extension beyond the abasic site. Taken together, these findings suggest that B35DNAP may perform faithful and processive genome replication in vivo and, when required, TLS of abasic sites.

scholarly journals Expression of the cancer-associated DNA polymerase ϵ P286R  in fission yeast leads to translesion synthesis polymerase dependent hypermutation and defective DNA replication

2021 ◽  
Author(s):  
Ignacio Soriano ◽  
Enrique Vazquez ◽  
Nagore de Leon ◽  
Sibyl Betrand ◽  
Ellen Heitzer ◽  
...  
Keyword(s):  
Fission Yeast ◽  
Dna Polymerase ◽  
Cytosine Methylation ◽  
Translesion Synthesis ◽  
Sequencing Analysis ◽  
Genes Encoding ◽  
Increased Sensitivity ◽  
Synthetically Lethal ◽  
Microsatellite Stability

Somatic mutations in the proofreading domain of the replicative DNA polymerase ϵ ( POLE- exonuclease domain mutations,  POLE -EDMs) are frequently found in colorectal and endometrial cancers and, occasionally, in other tumours. POLE-associated cancers typically display hypermutation, microsatellite stability and a unique mutational signature, with a predominance of C > A transversions in the context TCT. To understand better the contribution of hypermutagenesis to tumour development, we have modelled the most recurrent  POLE -EDM ( POLE-P286R ) in  Schizosaccharomyces pombe . Whole-genome sequencing analysis revealed that the corresponding  pol2-P287R  allele also has a strong mutator effect in vivo, with a high frequency of base substitutions and relatively few frameshift mutations. The mutations are equally distributed across different genomic regions, but they occur within an AT-rich context. The most abundant base-pair changes are T C T > T A T transversions and, in contrast to human mutations, T C G > T T G transitions are not elevated, likely due to the absence of cytosine methylation in fission yeast. The  pol2-P287R  variant has an increased sensitivity to elevated dNTP levels and DNA damaging agents, and shows reduced viability on depletion of the Pfh1 helicase. In addition, S phase is aberrant and RPA foci are elevated, suggestive of persistent ssDNA or DNA damage, and the  pol2-P287R  mutation is synthetically lethal with  rad3  inactivation ,  indicative of checkpoint activation. Significantly, deletion of genes encoding some translesion synthesis polymerases, most notably Pol κ, partially suppresses  pol2-P287R  hypermutation, indicating that polymerase switching contributes to this phenotype.

scholarly journals Expression of the cancer-associated DNA polymerase ε P286R in fission yeast leads to translesion synthesis polymerase dependent hypermutation and defective DNA replication

PLoS Genetics ◽  
2021 ◽  
Vol 17 (7) ◽  
pp. e1009526
Author(s):  
Ignacio Soriano ◽  
Enrique Vazquez ◽  
Nagore De Leon ◽  
Sibyl Bertrand ◽  
Ellen Heitzer ◽  
...  
Keyword(s):  
Fission Yeast ◽  
Dna Polymerase ◽  
Cytosine Methylation ◽  
Translesion Synthesis ◽  
Sequencing Analysis ◽  
Genes Encoding ◽  
Increased Sensitivity ◽  
Synthetically Lethal ◽  
Genomic Regions

Somatic and germline mutations in the proofreading domain of the replicative DNA polymerase ε (POLE-exonuclease domain mutations, POLE-EDMs) are frequently found in colorectal and endometrial cancers and, occasionally, in other tumours. POLE-associated cancers typically display hypermutation, and a unique mutational signature, with a predominance of C > A transversions in the context TCT and C > T transitions in the context TCG. To understand better the contribution of hypermutagenesis to tumour development, we have modelled the most recurrent POLE-EDM (POLE-P286R) in Schizosaccharomyces pombe. Whole-genome sequencing analysis revealed that the corresponding pol2-P287R allele also has a strong mutator effect in vivo, with a high frequency of base substitutions and relatively few indel mutations. The mutations are equally distributed across different genomic regions, but in the immediate vicinity there is an asymmetry in AT frequency. The most abundant base-pair changes are TCT > TAT transversions and, in contrast to human mutations, TCG > TTG transitions are not elevated, likely due to the absence of cytosine methylation in fission yeast. The pol2-P287R variant has an increased sensitivity to elevated dNTP levels and DNA damaging agents, and shows reduced viability on depletion of the Pfh1 helicase. In addition, S phase is aberrant and RPA foci are elevated, suggestive of ssDNA or DNA damage, and the pol2-P287R mutation is synthetically lethal with rad3 inactivation, indicative of checkpoint activation. Significantly, deletion of genes encoding some translesion synthesis polymerases, most notably Pol κ, partially suppresses pol2-P287R hypermutation, indicating that polymerase switching contributes to this phenotype.

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