Quantitative analysis of the mutagenic potential of 1-aminopyrene-DNA adduct bypass catalyzed by Y-family DNA polymerases

N2,3-Ethenoguanine (N2,3-ϵG) is one of the exocyclic DNA adducts produced by endogenous processes (e.g. lipid peroxidation) and exposure to bioactivated vinyl monomers such as vinyl chloride, which is a known human carcinogen. Existing studies exploring the miscoding potential of this lesion are quite indirect because of the lability of the glycosidic bond. We utilized a 2′-fluoro isostere approach to stabilize this lesion and synthesized oligonucleotides containing 2′-fluoro-N2,3-ϵ-2′-deoxyarabinoguanosine to investigate the miscoding potential of N2,3-ϵG by Y-family human DNA polymerases (pols). In primer extension assays, pol η and pol κ replicated through N2,3-ϵG, whereas pol ι and REV1 yielded only 1-base incorporation. Steady-state kinetics revealed that dCTP incorporation is preferred opposite N2,3-ϵG with relative efficiencies in the order of pol κ > REV1 > pol η ≈ pol ι, and dTTP misincorporation is the major miscoding event by all four Y-family human DNA pols. Pol ι had the highest dTTP misincorporation frequency (0.71) followed by pol η (0.63). REV1 misincorporated dTTP and dGTP with much lower frequencies. Crystal structures of pol ι with N2,3-ϵG paired to dCTP and dTTP revealed Hoogsteen-like base pairing mechanisms. Two hydrogen bonds were observed in the N2,3-ϵG:dCTP base pair, whereas only one appears to be present in the case of the N2,3-ϵG:dTTP pair. Base pairing mechanisms derived from the crystal structures explain the slightly favored dCTP insertion for pol ι in steady-state kinetic analysis. Taken together, these results provide a basis for the mutagenic potential of N2,3-ϵG.

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Miscoding Properties of 6α- and 6β-Diastereoisomers of theN2-(Estradiol-6-yl)-2′-deoxyguanosine DNA Adduct by Y-Family Human DNA Polymerases†

Biochemistry ◽

10.1021/bi7022255 ◽

2008 ◽

Vol 47 (25) ◽

pp. 6695-6701 ◽

Cited By ~ 7

Author(s):

Kinning Poon ◽

Shinji Itoh ◽

Naomi Suzuki ◽

Y. R. Santosh Laxmi ◽

Itsuo Yoshizawa ◽

...

Keyword(s):

Dna Polymerases ◽

Dna Adduct ◽

Human Dna ◽

Y Family

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Implications of inhibition of Rev1 interaction with Y family DNA polymerases for cisplatin chemotherapy

Genes & Development ◽

10.1101/gad.348662.121 ◽

2021 ◽

Author(s):

Jung-Hoon Yoon ◽

Robert E. Johnson ◽

Louise Prakash ◽

Satya Prakash

Keyword(s):

Adverse Effects ◽

Cancer Cells ◽

Dna Polymerases ◽

Translesion Synthesis ◽

Dna Adduct ◽

Secondary Malignancies ◽

Normal Cells ◽

Cross Links ◽

Y Family

Chemotherapy with cisplatin becomes limiting due to toxicity and secondary malignancies. In principle, therapeutics could be improved by targeting translesion synthesis (TLS) polymerases (Pols) that promote replication through intrastrand cross-links, the major cisplatin-induced DNA adduct. However, to specifically target malignancies with minimal adverse effects on normal cells, a good understanding of TLS mechanisms in normal versus cancer cells is paramount. We show that in normal cells, TLS through cisplatin intrastrand cross-links is promoted by Polη- or Polι-dependent pathways, both of which require Rev1 as a scaffolding component. In contrast, cancer cells require Rev1-Polζ. Our findings that a recently identified Rev1 inhibitor, JH-RE-06, purported to specifically disrupt Rev1 interaction with Polζ to block TLS through cisplatin adducts in cancer cells, abrogates Rev1's ability to function with Y family Pols as well, implying that by inactivating Rev1-dependent TLS in normal cells, this inhibitor will exacerbate the toxicity and tumorigenicity of chemotherapeutics with cisplatin.

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In VitroBypass of the Major Malondialdehyde- and Base Propenal-Derived DNA Adduct by Human Y-family DNA Polymerases κ, ι, and Rev1

Biochemistry ◽

10.1021/bi1009024 ◽

2010 ◽

Vol 49 (38) ◽

pp. 8415-8424 ◽

Cited By ~ 18

Author(s):

Leena Maddukuri ◽

Robert L. Eoff ◽

Jeong-Yun Choi ◽

Carmelo J. Rizzo ◽

F. Peter Guengerich ◽

...

Keyword(s):

Dna Polymerases ◽

Dna Adduct ◽

Y Family

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Mechanism of Translesion Synthesis Past an Equine Estrogen-DNA Adduct by Y-Family DNA Polymerases

Journal of Molecular Biology ◽

10.1016/j.jmb.2007.06.009 ◽

2007 ◽

Vol 371 (5) ◽

pp. 1151-1162 ◽

Cited By ~ 8

Author(s):

Manabu Yasui ◽

Naomi Suzuki ◽

Xiaoping Liu ◽

Yoshinori Okamoto ◽

Sung Yeon Kim ◽

...

Keyword(s):

Dna Polymerases ◽

Translesion Synthesis ◽

Dna Adduct ◽

Y Family

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Mutagenic Potential of 8-Oxo-7,8-dihydro-2′-deoxyguanosine Bypass Catalyzed by Human Y-Family DNA Polymerases

Chemical Research in Toxicology ◽

10.1021/tx500088e ◽

2014 ◽

Vol 27 (5) ◽

pp. 931-940 ◽

Cited By ~ 9

Author(s):

David J. Taggart ◽

Saul W. Fredrickson ◽

Varun V. Gadkari ◽

Zucai Suo

Keyword(s):

Dna Polymerases ◽

Mutagenic Potential ◽

Y Family

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Synthetic Nucleotides as Probes of DNA Polymerase Specificity

Journal of Nucleic Acids ◽

10.1155/2012/530963 ◽

2012 ◽

Vol 2012 ◽

pp. 1-17 ◽

Cited By ~ 10

Author(s):

Jason M. Walsh ◽

Penny J. Beuning

Keyword(s):

Dna Polymerase ◽

Dna Polymerases ◽

Polymerase Activity ◽

Chemical Diversity ◽

Dna Polymerase I ◽

Mutagenic Potential ◽

Natural Agents ◽

Polymerase I ◽

Y Family ◽

Insight Into

The genetic code is continuously expanding with new nucleobases designed to suit specific research needs. These synthetic nucleotides are used to study DNA polymerase dynamics and specificity and may even inhibit DNA polymerase activity. The availability of an increasing chemical diversity of nucleotides allows questions of utilization by different DNA polymerases to be addressed. Much of the work in this area deals with the A family DNA polymerases, for example,Escherichia coliDNA polymerase I, which are DNA polymerases involved in replication and whose fidelity is relatively high, but more recent work includes other families of polymerases, including the Y family, whose members are known to be error prone. This paper focuses on the ability of DNA polymerases to utilize nonnatural nucleotides in DNA templates or as the incoming nucleoside triphosphates. Beyond the utility of nonnatural nucleotides as probes of DNA polymerase specificity, such entities can also provide insight into the functions of DNA polymerases when encountering DNA that is damaged by natural agents. Thus, synthetic nucleotides provide insight into how polymerases deal with nonnatural nucleotides as well as into the mutagenic potential of nonnatural nucleotides.

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