Error-prone bypass of certain DNA lesions by the human DNA polymerase κ

The Escherichia coli protein DinB is a newly identified error-prone DNA polymerase. Recently, a human homolog of DinB was identified and named DINB1. We report that the DINB1gene encodes a DNA polymerase (designated polκ), which incorporates mismatched bases on a nondamaged template with a high frequency. Moreover, polκ bypasses an abasic site andN-2–acetylaminofluorene (AAF)-adduct in an error-prone manner but does not bypass a cis–syn or (6-4) thymine–thymine dimer or a cisplatin-adduct. Therefore, our results implicate an important role for polκ in the mutagenic bypass of certain types of DNA lesions.

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Expression and Activity of Human DNA Polymerase ^|^eta; in Escherichia coli

Genes and Environment ◽

10.3123/jemsge.35.10 ◽

2013 ◽

Vol 35 (1) ◽

pp. 10-20 ◽

Cited By ~ 1

Author(s):

Petr Gr^|^uacute;z ◽

Takehiko Nohmi

Keyword(s):

Escherichia Coli ◽

Dna Polymerase ◽

Polymerase Eta ◽

Human Dna ◽

Dna Polymerase Eta ◽

Expression And Activity

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Faculty Opinions recommendation of Mutations in human DNA polymerase eta motif II alter bypass of DNA lesions.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1003483.37805 ◽

2002 ◽

Author(s):

Errol C Friedberg

Keyword(s):

Dna Polymerase ◽

Dna Lesions ◽

Polymerase Eta ◽

Human Dna ◽

Dna Polymerase Eta

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Faculty Opinions recommendation of Analysis of translesion replication across an abasic site by DNA polymerase IV of Escherichia coli.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1016604.201051 ◽

2003 ◽

Author(s):

Errol C Friedberg

Keyword(s):

Escherichia Coli ◽

Dna Polymerase ◽

Abasic Site ◽

Dna Polymerase Iv

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Human DNA Polymerase ι Promotes Replication through a Ring-Closed Minor-Groove Adduct That Adopts a syn Conformation in DNA

Molecular and Cellular Biology ◽

10.1128/mcb.25.19.8748-8754.2005 ◽

2005 ◽

Vol 25 (19) ◽

pp. 8748-8754 ◽

Cited By ~ 30

Author(s):

William T. Wolfle ◽

Robert E. Johnson ◽

Irina G. Minko ◽

R. Stephen Lloyd ◽

Satya Prakash ◽

...

Keyword(s):

Dna Polymerase ◽

Dna Polymerases ◽

Minor Groove ◽

Dna Lesions ◽

Unsaturated Aldehyde ◽

Cyclic Form ◽

Human Dna ◽

A Minor ◽

Dna Polymerase Ι

ABSTRACT Acrolein, an α,β-unsaturated aldehyde, is generated in vivo as the end product of lipid peroxidation and from oxidation of polyamines. The reaction of acrolein with the N 2 group of guanine in DNA leads to the formation of a cyclic adduct, γ-hydroxy-1,N 2-propano-2′-deoxyguanosine (γ-HOPdG). Previously, we have shown that proficient replication through the γ-HOPdG adduct can be mediated by the sequential action of human DNA polymerases (Pols) ι and κ, in which Polι incorporates either pyrimidine opposite γ-HOPdG, but Polκ extends only from the cytosine. Since γ-HOPdG can adopt either a ring-closed cyclic form or a ring-opened form in DNA, to better understand the mechanisms that Pols ι and κ employ to promote replication through this lesion, we have examined the ability of these polymerases to replicate through the structural analogs of γ-HOPdG that are permanently either ring closed or ring opened. Our studies with these model adducts show that whereas the ring-opened form of γ-HOPdG is not inhibitory to synthesis by human Pols η, ι, or κ, only Polι is able to incorporate nucleotides opposite the ring-closed form, which is known to adopt a syn conformation in DNA. From these studies, we infer that (i) Pols η, ι, and κ have the ability to proficiently replicate through minor-groove DNA lesions that do not perturb the Watson-Crick hydrogen bonding of the template base with the incoming nucleotide, and (ii) Polι can accommodate a minor-groove-adducted template purine which adopts a syn conformation in DNA and forms a Hoogsteen base pair with the incoming nucleotide.

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Structural insights into the promutagenic bypass of the major cisplatin-induced DNA lesion

Biochemical Journal ◽

10.1042/bcj20190906 ◽

2020 ◽

Vol 477 (5) ◽

pp. 937-951

Author(s):

Hala Ouzon-Shubeita ◽

Caroline K. Vilas ◽

Seongmin Lee

Keyword(s):

Dna Polymerase ◽

Active Site ◽

Cisplatin Resistance ◽

Structural Basis ◽

Abasic Site ◽

Dna Polymerase Β ◽

Dna Lesion ◽

Human Dna ◽

Optimal Conformation ◽

Structural Insights

The cisplatin-1,2-d(GpG) (Pt-GG) intrastrand cross-link is the predominant DNA lesion generated by cisplatin. Cisplatin has been shown to predominantly induce G to T mutations and Pt-GG permits significant misincorporation of dATP by human DNA polymerase β (polβ). In agreement, polβ overexpression, which is frequently observed in cancer cells, is linked to cisplatin resistance and a mutator phenotype. However, the structural basis for the misincorporation of dATP opposite Pt-GG is unknown. Here, we report the first structures of a DNA polymerase inaccurately bypassing Pt-GG. We solved two structures of polβ misincorporating dATP opposite the 5′-dG of Pt-GG in the presence of Mg2+ or Mn2+. The Mg2+-bound structure exhibits a sub-optimal conformation for catalysis, while the Mn2+-bound structure is in a catalytically more favorable semi-closed conformation. In both structures, dATP does not form a coplanar base pairing with Pt-GG. In the polβ active site, the syn-dATP opposite Pt-GG appears to be stabilized by protein templating and pi stacking interactions, which resembles the polβ-mediated dATP incorporation opposite an abasic site. Overall, our results suggest that the templating Pt-GG in the polβ active site behaves like an abasic site, promoting the insertion of dATP in a non-instructional manner.

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Preferential Incorporation of G Opposite Template T by the Low-Fidelity Human DNA Polymerase ι

Molecular and Cellular Biology ◽

10.1128/mcb.20.19.7099-7108.2000 ◽

2000 ◽

Vol 20 (19) ◽

pp. 7099-7108 ◽

Cited By ~ 137

Author(s):

Yanbin Zhang ◽

Fenghua Yuan ◽

Xiaohua Wu ◽

Zhigang Wang

Keyword(s):

Dna Synthesis ◽

Dna Polymerase ◽

Polymerase Activity ◽

Abasic Site ◽

Base Pairing ◽

Base Pairs ◽

Dna Lesion ◽

Biological Source ◽

Human Dna ◽

Base Selection

ABSTRACT DNA polymerase activity is essential for replication, recombination, repair, and mutagenesis. All DNA polymerases studied so far from any biological source synthesize DNA by the Watson-Crick base-pairing rule, incorporating A, G, C, and T opposite the templates T, C, G, and A, respectively. Non-Watson-Crick base pairs would lead to mutations. In this report, we describe the ninth human DNA polymerase, Polι, encoded by the RAD30B gene. We show that human Polι violates the Watson-Crick base-pairing rule opposite template T. During base selection, human Polι preferred T-G base pairing, leading to G incorporation opposite template T. The resulting T-G base pair was less efficiently extended by human Polι compared to the Watson-Crick base pairs. Consequently, DNA synthesis frequently aborted opposite template T, a property we designated the T stop. This T stop restricted human Polι to a very short stretch of DNA synthesis. Furthermore, kinetic analyses show that human Polι copies template C with extraordinarily low fidelity, misincorporating T, A, and C with unprecedented frequencies of 1/9, 1/10, and 1/11, respectively. Human Polι incorporated one nucleotide opposite a template abasic site more efficiently than opposite a template T, suggesting a role for human Polι in DNA lesion bypass. The unique features of preferential G incorporation opposite template T and T stop suggest that DNA Polι may additionally play a specialized function in human biology.

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