Multiple PolK (POLK) transcripts in mammalian testis

Guo, Caixia; Gao, Tianshu; Confer, Nils; Velasco-Miguel, Susana; Friedberg, Errol C.

doi:10.1016/j.dnarep.2004.10.006

Cited by 9 publications

(8 citation statements)

References 24 publications

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“…To determine whether the Polk derivative harboring D197A-E198A is expressed at levels comparable to that of the wild-type Polk, we conducted Western blotting analyses. Bands corresponding to the expected molecular size of mouse Polk (96 kDa) were detected and the band pattern of proteins was similar to that in a previous report [50]. The similar intensity in the thymus, adrenal cortex, liver, and testis were detected between the targeted and the wild-type mice (Fig.…”

Section: Generation Of Inactivated Polk Ki Micesupporting

confidence: 87%

In vivo evidence that DNA polymerase kappa is responsible for error-free bypass across DNA cross-links induced by mitomycin C

Takeiri¹,

Wada²,

Motoyama³

et al. 2014

DNA Repair

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Section: Generation Of Inactivated Polk Ki Micesupporting

confidence: 87%

In vivo evidence that DNA polymerase kappa is responsible for error-free bypass across DNA cross-links induced by mitomycin C

Takeiri¹,

Wada²,

Motoyama³

et al. 2014

DNA Repair

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“…We reported previously the presence of multiple PolK transcripts in mouse testis (27). Many of the putative Pol protein isoforms thus identified lack UBZ domains.…”

Section: Discussionmentioning

confidence: 79%

“…Anti-FLAG M2 agarose affinity gel and anti-FLAG M2 monoclonal antibodies were purchased from Sigma. Hamster polyclonal antiserum against mouse Pol was made by our laboratory (27). Rabbit polyclonal antiserum against mouse Pol was generated with a 14-amino acid peptide (CNYLKIDTPRQE-ANE) containing an N-terminal cysteine residue conjugated with keyhole limpet hemocyanin as described (28).…”

Section: Methodsmentioning

confidence: 99%

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Requirements for the Interaction of Mouse Polκ with Ubiquitin and Its Biological Significance

Guo

Tang

Bienko

et al. 2008

Journal of Biological Chemistry

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Pol protein is a eukaryotic member of the DinB/Pol 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 Pol in this process has remained unknown. Here we report that mouse Pol can physically interact with ubiquitin by yeast two-hybrid screening, glutathione S-transferase pulldown, and immunoprecipitation methods. The association of Pol with ubiquitin requires the ubiquitin-binding motifs located at the C terminus of Pol. In addition, Pol 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 Pol. The ubiquitin-binding motifs are also required for Pol to form nuclear foci after UV radiation. However, the ubiquitin-binding motifs do not affect Pol half-life. Finally, we have examined levels of Pol expression following the exposure of mouse cells to benzo[a]pyrene-dihydrodiol epoxide or UVB radiation. Translesion DNA synthesis (TLS)3 is one of several biochemical mechanisms by which cells can tolerate DNA damage that arrests semiconservative DNA synthesis (1, 2). This process requires the action of specialized DNA polymerases present in bacteria (such as Escherichia coli), lower eukaryotes, and vertebrates. Lower eukaryotes, particularly vertibrates, contain multiple such enzymes, suggesting the ability to bypass many types of DNA damage.Several specialized DNA polymerases are members of a novel polymerase family, the Y-family (3). These enzymes are devoid of 3Ј 3 5Ј proofreading exonuclease activity and replicate undamaged DNA in vitro with low fidelity and weak processivity (4). Members of this family in mammalian cells include Pol, Pol, and Pol, all of which can extend primers for varying distances past various types of template damage (4). A fourth member of the Y-family, REV1 protein, is able to catalyze the incorporation of only one or two dCMP moieties, regardless of the template base composition (5). Pol, Pol, and Pol have been shown to interact with REV1 protein via a highly conserved C-terminal domain in REV1 (6 -8). These polymerases also interact with PCNA (9, 10), and recent observations suggest that PCNA plays a key role in promoting the access of specialized polymerases to arrested replication forks (11-15).Disruption of the PolK gene in mouse and chicken cells results in significant sensitivity to killing by benzo[a]pyrenedihydrodiol epoxide (BPDE) and UV radiation (16 -18). Poldeficient mouse embryonic stem and fibroblast cells also show moderate sensitivity to methyl methanesulfonate (19). Consistent with these results, primer extension assays have shown that human Pol can support TLS across sites of base loss, acetylaminofluorene-G adducts, benzo[a]pyrene-G adducts, and thymine glycol (4). However, the enzyme does not support primer extension past thymine-thymine (TϽϾT) dimers or [6,4]pyrimidine-pyrimidone p...

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“…Mouse Polk is ubiquitously expressed, with highest expression levels in testis [111,137,138]. Multiple transcripts are present in this tissue [139] and Polk expression is confined to meiotic spermatocytes and postmeiotic spermatids, suggesting a specific role of Polj in spermatogenesis [138]. Polk mRNA is also highly expressed in the adrenal cortex in mouse embryos and adult animals [138].…”

Section: Polk Gene Expression and Regulationmentioning

confidence: 99%

Y-family DNA polymerases in mammalian cells

Guo

Kosarek-Stancel

Tang

et al. 2009

Cell. Mol. Life Sci.

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130

159

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Eukaryotic genomes are replicated with high fidelity to assure the faithful transmission of genetic information from one generation to the next. The accuracy of replication relies heavily on the ability of replicative DNA polymerases to efficiently select correct nucleotides for the polymerization reaction and, using their intrinsic exonuclease activities, to excise mistakenly incorporated nucleotides. Cells also possess a variety of specialized DNA polymerases that, by a process called translesion DNA synthesis (TLS), help overcome replication blocks when unrepaired DNA lesions stall the replication machinery. This review considers the properties of the Y-family (a subset of specialized DNA polymerases) and their roles in modulating spontaneous and genotoxic-induced mutations in mammals. We also review recent insights into the molecular mechanisms that regulate PCNA monoubiquitination and DNA polymerase switching during TLS and discuss the potential of using Y-family DNA polymerases as novel targets for cancer prevention and therapy.

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Multiple PolK (POLK) transcripts in mammalian testis

Cited by 9 publications

References 24 publications

In vivo evidence that DNA polymerase kappa is responsible for error-free bypass across DNA cross-links induced by mitomycin C

In vivo evidence that DNA polymerase kappa is responsible for error-free bypass across DNA cross-links induced by mitomycin C

Requirements for the Interaction of Mouse Polκ with Ubiquitin and Its Biological Significance

Y-family DNA polymerases in mammalian cells

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