A Comprehensive Strategy to Discover Inhibitors of the Translesion Synthesis DNA Polymerase κ

Kinrin Yamanaka, Dorjbal Dorjsuren, Robert L. Eoff, Martin Egli, David J. Maloney, Ajit Jadhav, Anton Simeonov, Robert (Stephen) Lloyd

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Human DNA polymerase kappa (pol κ) is a translesion synthesis (TLS) polymerase that catalyzes TLS past various minor groove lesions including N2-dG linked acrolein- and polycyclic aromatic hydrocarbon-derived adducts, as well as N2-dG DNA-DNA interstrand cross-links introduced by the chemotherapeutic agent mitomycin C. It also processes ultraviolet light-induced DNA lesions. Since pol κ TLS activity can reduce the cellular toxicity of chemotherapeutic agents and since gliomas overexpress pol κ, small molecule library screens targeting pol κ were conducted to initiate the first step in the development of new adjunct cancer therapeutics. A high-throughput, fluorescence-based DNA strand displacement assay was utilized to screen ~16,000 bioactive compounds, and the 60 top hits were validated by primer extension assays using non-damaged DNAs. Candesartan cilexetil, manoalide, and MK-886 were selected as proof-of-principle compounds and further characterized for their specificity toward pol κ by primer extension assays using DNAs containing a site-specific acrolein-derived, ring-opened reduced form of γ-HOPdG. Furthermore, candesartan cilexetil could enhance ultraviolet light-induced cytotoxicity in xeroderma pigmentosum variant cells, suggesting its inhibitory effect against intracellular pol κ. In summary, this investigation represents the first high-throughput screening designed to identify inhibitors of pol κ, with the characterization of biochemical and biologically relevant endpoints as a consequence of pol κ inhibition. These approaches lay the foundation for the future discovery of compounds that can be applied to combination chemotherapy.

Original languageEnglish (US)
Article numbere45032
JournalPLoS One
Volume7
Issue number10
DOIs
StatePublished - Oct 8 2012

Fingerprint

DNA-directed DNA polymerase
DNA-Directed DNA Polymerase
synthesis
DNA
acrolein
Acrolein
Assays
Ultraviolet Rays
L 663536
lesions (animal)
ultraviolet radiation
cytotoxicity
assays
Throughput
Small Molecule Libraries
mitomycin
Chemotherapy
Polycyclic Aromatic Hydrocarbons
Mitomycin
polycyclic aromatic hydrocarbons

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

A Comprehensive Strategy to Discover Inhibitors of the Translesion Synthesis DNA Polymerase κ. / Yamanaka, Kinrin; Dorjsuren, Dorjbal; Eoff, Robert L.; Egli, Martin; Maloney, David J.; Jadhav, Ajit; Simeonov, Anton; Lloyd, Robert (Stephen).

In: PLoS One, Vol. 7, No. 10, e45032, 08.10.2012.

Research output: Contribution to journalArticle

Yamanaka, K, Dorjsuren, D, Eoff, RL, Egli, M, Maloney, DJ, Jadhav, A, Simeonov, A & Lloyd, RS 2012, 'A Comprehensive Strategy to Discover Inhibitors of the Translesion Synthesis DNA Polymerase κ', PLoS One, vol. 7, no. 10, e45032. https://doi.org/10.1371/journal.pone.0045032
Yamanaka K, Dorjsuren D, Eoff RL, Egli M, Maloney DJ, Jadhav A et al. A Comprehensive Strategy to Discover Inhibitors of the Translesion Synthesis DNA Polymerase κ. PLoS One. 2012 Oct 8;7(10). e45032. https://doi.org/10.1371/journal.pone.0045032
Yamanaka, Kinrin ; Dorjsuren, Dorjbal ; Eoff, Robert L. ; Egli, Martin ; Maloney, David J. ; Jadhav, Ajit ; Simeonov, Anton ; Lloyd, Robert (Stephen). / A Comprehensive Strategy to Discover Inhibitors of the Translesion Synthesis DNA Polymerase κ. In: PLoS One. 2012 ; Vol. 7, No. 10.
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