Requirements for the interaction of mouse Polκ with ubiquitin and its biological significance

Caixia Guo, Tie Shan Tang, Marzena Bienko, Ivan Dikic, Errol C. Friedberg

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Polκ protein is a eukaryotic member of the DinB/Polκ branch of the Y-family DNA polymerases, which are involved in the tolerance of DNA damage by replicative bypass. Despite universal conservation through evolution, the precise role(s) of Polκ in this process has remained unknown. Here we report that mouse Polκ can physically interact with ubiquitin by yeast two-hybrid screening, glutathione S-transferase pulldown, and immunoprecipitation methods. The association of Polκ with ubiquitin requires the ubiquitin-binding motifs located at the C terminus of Polκ. In addition, Polκ binds with monoubiquitinated proliferating cell nuclear antigen (PCNA) more robustly than with non-ubiquitinated PCNA. The ubiquitin-binding motifs mediate the enhanced association between monoubiquitinated PCNA and Polκ. The ubiquitin-binding motifs are also required for Polκ to form nuclear foci after UV radiation. However, the ubiquitin-binding motifs do not affect Polκ half-life. Finally, we have examined levels of Polκ expression following the exposure of mouse cells to benzo[a]pyrene-dihydrodiol epoxide or UVB radiation.

Original languageEnglish (US)
Pages (from-to)4658-4664
Number of pages7
JournalJournal of Biological Chemistry
Volume283
Issue number8
DOIs
StatePublished - Feb 22 2008

Fingerprint

Ubiquitin
Proliferating Cell Nuclear Antigen
pol Gene Products
Radiation
Benzo(a)pyrene
Epoxy Compounds
DNA-Directed DNA Polymerase
Glutathione Transferase
Immunoprecipitation
Ultraviolet radiation
Yeast
DNA Damage
Half-Life
Conservation
Screening
Yeasts
DNA

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Requirements for the interaction of mouse Polκ with ubiquitin and its biological significance. / Guo, Caixia; Tang, Tie Shan; Bienko, Marzena; Dikic, Ivan; Friedberg, Errol C.

In: Journal of Biological Chemistry, Vol. 283, No. 8, 22.02.2008, p. 4658-4664.

Research output: Contribution to journalArticle

Guo, Caixia ; Tang, Tie Shan ; Bienko, Marzena ; Dikic, Ivan ; Friedberg, Errol C. / Requirements for the interaction of mouse Polκ with ubiquitin and its biological significance. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 8. pp. 4658-4664.
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