scholarly journals Role of DNA Polymerase η in the Bypass of a (6-4) TT Photoproduct

2001 ◽  
Vol 21 (10) ◽  
pp. 3558-3563 ◽  
Author(s):  
Robert E. Johnson ◽  
Lajos Haracska ◽  
Satya Prakash ◽  
Louise Prakash
Keyword(s):  
Dna Polymerase ◽  
Xeroderma Pigmentosum ◽  
Uv Light ◽  
Dna Lesions ◽  
Variant Form ◽  
Replication Machinery ◽  
Genetic Studies ◽  
Induced Mutagenesis ◽  
Combined Action

ABSTRACT UV light-induced DNA lesions block the normal replication machinery. Eukaryotic cells possess DNA polymerase η (Polη), which has the ability to replicate past a cis-syn thymine-thymine (TT) dimer efficiently and accurately, and mutations in human Polη result in the cancer-prone syndrome, the variant form of xeroderma pigmentosum. Here, we test Polη for its ability to bypass a (6-4) TT lesion which distorts the DNA helix to a much greater extent than acis-syn TT dimer. Opposite the 3′ T of a (6-4) TT photoproduct, both yeast and human Polη preferentially insert a G residue, but they are unable to extend from the inserted nucleotide. DNA Polζ, essential for UV induced mutagenesis, efficiently extends from the G residue inserted opposite the 3′ T of the (6-4) TT lesion by Polη, and Polζ inserts the correct nucleotide A opposite the 5′ T of the lesion. Thus, the efficient bypass of the (6-4) TT photoproduct is achieved by the combined action of Polη and Polζ, wherein Polη inserts a nucleotide opposite the 3′ T of the lesion and Polζ extends from it. These biochemical observations are in concert with genetic studies in yeast indicating that mutations occur predominantly at the 3′ T of the (6-4) TT photoproduct and that these mutations frequently exhibit a 3′ T→C change that would result from the insertion of a G opposite the 3′ T of the (6-4) TT lesion.

scholarly journals Requirement of DNA Polymerase η for Error-Free Bypass of UV-Induced CC and TC Photoproducts

2001 ◽  
Vol 21 (1) ◽  
pp. 185-188 ◽  
Author(s):  
Sung-Lim Yu ◽  
Robert E. Johnson ◽  
Satya Prakash ◽  
Louise Prakash
Keyword(s):  
Dna Polymerase ◽  
Uv Light ◽  
Wide Spectrum ◽  
Variant Form ◽  
Induced Mutations ◽  
Uv Mutagenesis ◽  
Genetic Studies ◽  
Human Dna ◽  
Skin Cancers ◽  
Cyclobutane Dimers

ABSTRACT The yeast RAD30-encoded DNA polymerase η (Polη) bypasses a cis-syn thymine-thymine dimer efficiently and accurately. Human DNA polymerase η functions similarly in the bypass of this lesion, and mutations in human Polη result in the cancer prone syndrome, the variant form of xeroderma pigmentosum. UV light, however, also elicits the formation ofcis-syn cyclobutane dimers and (6-4) photoproducts at 5′-CC-3′ and 5′-TC-3′ sites, and in both yeast and human DNA, UV-induced mutations occur primarily by 3′ C to T transitions. Genetic studies presented here reveal a role for yeast Polη in the error-free bypass of cyclobutane dimers and (6-4) photoproducts formed at CC and TC sites. Thus, by preventing UV mutagenesis at a wide spectrum of dipyrimidine sites, Polη plays a pivotal role in minimizing the incidence of sunlight-induced skin cancers in humans.

scholarly journals Yeast DNA Polymerase ζ Is an Efficient Extender of Primer Ends Opposite from 7,8-Dihydro-8-Oxoguanine and O6-Methylguanine

2003 ◽  
Vol 23 (4) ◽  
pp. 1453-1459 ◽  
Author(s):  
Lajos Haracska ◽  
Satya Prakash ◽  
Louise Prakash
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Polymerase ◽  
Uv Light ◽  
Dna Lesions ◽  
Geometric Distortion ◽  
Cyclobutane Pyrimidine Dimers ◽  
Genetic Studies ◽  
Abasic Sites ◽  
Dna Damaging Agents ◽  
Pyrimidine Dimers

ABSTRACT Genetic studies in Saccharomyces cerevisiae have indicated the requirement of DNA polymerase (Pol) ζ for mutagenesis induced by UV light and by other DNA damaging agents. However, on its own, Polζ is highly inefficient at replicating through DNA lesions; rather, it promotes their mutagenic bypass by extending from the nucleotide inserted opposite the lesion by another DNA polymerase. So far, such a role for Polζ has been established for cyclobutane pyrimidine dimers, (6-4) dipyrimidine photoproducts, and abasic sites. Here, we examine whether Polζ can replicate through the 7,8-dihydro-8-oxoguanine (8-oxoG) and O 6-methylguanine (m6G) lesions. We chose these two lesions for this study because the replicative polymerase, Polδ, can replicate through them, albeit weakly. We found that Polζ is very inefficient at inserting nucleotides opposite both these lesions, but it can efficiently extend from the nucleotides inserted opposite them by Polδ. Also, the most efficient bypass of 8-oxoG and m6G lesions occurs when Polδ is combined with Polζ, indicating a role for Polζ in extending from the nucleotides inserted opposite these lesions by Polδ. Thus, Polζ is a highly specialized polymerase that can proficiently extend from the primer ends opposite DNA lesions, irrespective of their degree of geometric distortion. Polζ, however, is unusually sensitive to geometric distortion of the templating residue, as it is highly inefficient at incorporating nucleotides even opposite the moderately distorting 8-oxoG and m6G lesions.

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

2003 ◽  
Vol 50 (4) ◽  
pp. 921-939 ◽  
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.

scholarly journals Effect of SOS-induced Pol II, Pol IV, and Pol V DNA polymerases on UV-induced mutagenesis and MFD repair in Escherichia coli cells.

2005 ◽  
Vol 52 (1) ◽  
pp. 139-147
Author(s):  
Michał Wrzesiński ◽  
Anetta Nowosielska ◽  
Jadwiga Nieminuszczy ◽  
Elzbieta Grzesiuk
Keyword(s):  
Escherichia Coli ◽  
Uv Light ◽  
Dna Polymerases ◽  
Dna Lesions ◽  
Uv Mutagenesis ◽  
Pol Ii ◽  
Pol Iv ◽  
Pol V ◽  
Induced Mutagenesis ◽  
Pol Iii

Irradiation of organisms with UV light produces genotoxic and mutagenic lesions in DNA. Replication through these lesions (translesion DNA synthesis, TSL) in Escherichia coli requires polymerase V (Pol V) and polymerase III (Pol III) holoenzyme. However, some evidence indicates that in the absence of Pol V, and with Pol III inactivated in its proofreading activity by the mutD5 mutation, efficient TSL takes place. The aim of this work was to estimate the involvement of SOS-inducible DNA polymerases, Pol II, Pol IV and Pol V, in UV mutagenesis and in mutation frequency decline (MFD), a mechanism of repair of UV-induced damage to DNA under conditions of arrested protein synthesis. Using the argE3-->Arg(+) reversion to prototrophy system in E. coli AB1157, we found that the umuDC-encoded Pol V is the only SOS-inducible polymerase required for UV mutagenesis, since in its absence the level of Arg(+) revertants is extremely low and independent of Pol II and/or Pol IV. The low level of UV-induced Arg(+) revertants observed in the AB1157mutD5DumuDC strain indicates that under conditions of disturbed proofreading activity of Pol III and lack of Pol V, UV-induced lesions are bypassed without inducing mutations. The presented results also indicate that Pol V may provide substrates for MFD repair; moreover, we suggest that only those DNA lesions which result from umuDC-directed UV mutagenesis are subject to MFD repair.

scholarly journals Rev1 promotes replication through UV lesions in conjunction with DNA polymerases η, ι, and κ but not DNA polymerase ζ

2015 ◽  
Vol 29 (24) ◽  
pp. 2588-2602 ◽  
Author(s):  
Jung-Hoon Yoon ◽  
Jeseong Park ◽  
Juan Conde ◽  
Maki Wakamiya ◽  
Louise Prakash ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Crucial Role ◽  
Genome Stability ◽  
Dna Polymerases ◽  
Translesion Synthesis ◽  
Dna Lesions ◽  
Human Cells ◽  
Uv Lesions ◽  
Human And Mouse

Translesion synthesis (TLS) DNA polymerases (Pols) promote replication through DNA lesions; however, little is known about the protein factors that affect their function in human cells. In yeast, Rev1 plays a noncatalytic role as an indispensable component of Polζ, and Polζ together with Rev1 mediates a highly mutagenic mode of TLS. However, how Rev1 functions in TLS and mutagenesis in human cells has remained unclear. Here we determined the role of Rev1 in TLS opposite UV lesions in human and mouse fibroblasts and showed that Rev1 is indispensable for TLS mediated by Polη, Polι, and Polκ but is not required for TLS by Polζ. In contrast to its role in mutagenic TLS in yeast, Rev1 promotes predominantly error-free TLS opposite UV lesions in humans. The identification of Rev1 as an indispensable scaffolding component for Polη, Polι, and Polκ, which function in TLS in highly specialized ways opposite a diverse array of DNA lesions and act in a predominantly error-free manner, implicates a crucial role for Rev1 in the maintenance of genome stability in humans.

scholarly journals DNA Processing Proteins Involved in the UV-Induced Stress Response of Sulfolobales

2015 ◽  
Vol 197 (18) ◽  
pp. 2941-2951 ◽  
Author(s):  
Marleen van Wolferen ◽  
Xiaoqing Ma ◽  
Sonja-Verena Albers
Keyword(s):  
Homologous Recombination ◽  
Uv Light ◽  
Dna Lesions ◽  
Dna Transfer ◽  
Content Type ◽  
Induced Stress ◽  
Cellular Aggregation ◽  
Uv Stress ◽  
Dna Processing

ABSTRACTTheupsoperon ofSulfolobusspecies is highly induced upon UV stress. Previous studies showed that the pili encoded by this operon are involved in cellular aggregation, which is essential for subsequent DNA exchange between cells, resulting in homologous recombination. The presence of this pilus system increases the fitness ofSulfolobuscells under UV light-induced stress conditions, as the transfer of DNA takes place in order to repair UV-induced DNA lesions via homologous recombination. Four conserved genes (saci_1497tosaci_1500) which encode proteins with putative DNA processing functions are present downstream of theupsoperon. In this study, we show that after UV treatment the cellular aggregation of strains withsaci_1497,saci_1498, andsaci_1500deletions is similar to that of wild-type strains; their survival rates, however, were reduced and similar to or lower than those of the pilus deletion strains, which could not aggregate anymore. DNA recombination assays indicated thatsaci_1498, encoding a ParB-like protein, plays an important role in DNA transfer. Moreover, biochemical analysis showed that the endonuclease III encoded bysaci_1497nicks UV-damaged DNA. In addition, RecQ-like helicase Saci_1500 is able to unwind homologous recombination intermediates, such as Holliday junctions. Interestingly, asaci_1500deletion mutant was more sensitive to UV light but not to the replication-stalling agents hydroxyurea and methyl methanesulfonate, suggesting that Saci_1500 functions specifically in the UV damage pathway. Together these results suggest a role of Saci_1497 to Saci_1500 in the repair or transfer of DNA that takes place after UV-induced damage to the genomic DNA ofSulfolobus acidocaldarius.IMPORTANCESulfolobalesspecies increase their fitness after UV stress by a UV-inducible pilus system that enables high rates of DNA exchange between cells. Downstream of the pilus operon, three genes that seem to play a role in the repair or transfer of the DNA betweenSulfolobuscells were identified, and their possible functions are discussed. Next to the previously described role of UV-inducible pili in the exchange of DNA, we have thereby increased our knowledge of DNA transfer at the level of DNA processing. This paper therefore contributes to the overall understanding of the DNA exchange mechanism amongSulfolobalescells.

scholarly journals Complex Formation with Rev1 Enhances the Proficiency of Saccharomyces cerevisiae DNA Polymerase ζ for Mismatch Extension and for Extension Opposite from DNA Lesions

2006 ◽  
Vol 26 (24) ◽  
pp. 9555-9563 ◽  
Author(s):  
Narottam Acharya ◽  
Robert E. Johnson ◽  
Satya Prakash ◽  
Louise Prakash
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Polymerase ◽  
Dna Lesions ◽  
Synthetic Activity ◽  
Lesion Bypass ◽  
Dna Lesion ◽  
C Terminus ◽  
Accessory Subunit ◽  
Induced Mutagenesis ◽  
Y Family

ABSTRACT Rev1, a Y family DNA polymerase (Pol) functions together with Polζ, a B family Pol comprised of the Rev3 catalytic subunit and Rev7 accessory subunit, in promoting translesion DNA synthesis (TLS). Extensive genetic studies with Saccharomyces cerevisiae have indicated a requirement of both Polζ and Rev1 for damage-induced mutagenesis, implicating their involvement in mutagenic TLS. Polζ is specifically adapted to promote the extension step of lesion bypass, as it proficiently extends primer termini opposite DNA lesions, and it is also a proficient extender of mismatched primer termini on undamaged DNAs. Since TLS through UV-induced lesions and various other DNA lesions does not depend upon the DNA-synthetic activity of Rev1, Rev1 must contribute to Polζ-dependent TLS in a nonenzymatic way. Here, we provide evidence for the physical association of Rev1 with Polζ and show that this binding is mediated through the C terminus of Rev1 and the polymerase domain of Rev3. Importantly, a rev1 mutant that lacks the C-terminal 72 residues which inactivate interaction with Rev3 exhibits the same high degree of UV sensitivity and defectiveness in UV-induced mutagenesis as that conferred by the rev1Δ mutation. We propose that Rev1 binding to Polζ is indispensable for the targeting of Polζ to the replication fork stalled at a DNA lesion. In addition to this structural role, Rev1 binding enhances the proficiency of Polζ for the extension of mismatched primer termini on undamaged DNAs and for the extension of primer termini opposite DNA lesions.

scholarly journals Deaminated purine bypass by DNA polymerase η

2021 ◽  
Vol 478 (7) ◽  
pp. 1309-1313
Author(s):  
Antonina Andreeva
Keyword(s):  
Crystal Structures ◽  
Recent Work ◽  
Dna Polymerase ◽  
Dna Lesions ◽  
Purine Bases

A recent work by Jung and colleagues (Biochem J.477, 4797–4810) provides an explanation of how DNA polymerase η replicates through deaminated purine bases such as xanthine and hypoxanthine. This commentary discusses the crystal structures of the polymerase η complexes that implicate the role of tautomerism in the bypass of these DNA lesions.

scholarly journals Role of AtPolζ, AtRev1, and AtPolη in UV Light-Induced Mutagenesis in Arabidopsis

2010 ◽  
Vol 155 (1) ◽  
pp. 414-420 ◽  
Author(s):  
Mayu Nakagawa ◽  
Shinya Takahashi ◽  
Atsushi Tanaka ◽  
Issay Narumi ◽  
Ayako N. Sakamoto
Keyword(s):  
Uv Light ◽  
Induced Mutagenesis

scholarly journals Role of Acinetobacter baumannii UmuD Homologs in Antibiotic Resistance Acquired through DNA Damage-Induced Mutagenesis

2013 ◽  
Vol 58 (3) ◽  
pp. 1771-1773 ◽  
Author(s):  
Jesús Aranda ◽  
Mario López ◽  
Enoy Leiva ◽  
Andrés Magán ◽  
Ben Adler ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Acinetobacter Baumannii ◽  
Dna Polymerase ◽  
Dna Polymerases ◽  
Wild Type ◽  
Content Type ◽  
Dna Polymerase V ◽  
Induced Mutagenesis ◽  
Related Survival

ABSTRACTThe role ofAcinetobacter baumanniiATCC 17978 UmuDC homologs A1S_0636-A1S_0637, A1S_1174-A1S_1173, and A1S_1389 (UmuDAb) in antibiotic resistance acquired through UV-induced mutagenesis was evaluated. Neither the growth rate nor the UV-related survival of any of the three mutants was significantly different from that of the wild-type parental strain. However, all mutants, and especially theumuDAbmutant, were less able to acquire resistance to rifampin and streptomycin through the activities of their error-prone DNA polymerases. Furthermore, in theA. baumanniimutant defective in theumuDAbgene, the spectrum of mutations included a dramatic reduction in the frequency of transition mutations, the mutagenic signature of the DNA polymerase V encoded byumuDC.

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