Reconstitution of DNA base excision-repair with purified human proteins: interaction between DNA polymerase beta and the XRCC1 protein.

To evaluate contribution of polymorphisms of the XRCC1 gene to the risk of colorectal cancer, we conducted a case-control study of 209 colorectal cancer cases and 209 age-and gender-matched controls in the Korean population. We tested the hypothesis by constructing allele combinations with known SNP. Allelic variants of the XRCC1 gene at codons 194, 280 and 399 were analyzed in lymphocyte DNA by PCR-RFLP. We observed an increased risk of colorectal cancer associated with the 399Gln allele. The odds ratio (OR) was 1.61 (95% confidence interval [CI] 1.09-2.39) for the 399Gln allele. When combined allele-specific OR were calculated after estimating frequencies, 3 common allele combinations were found to be associated with an increased risk of colorectal cancer. The OR for the 194Trp-280Arg-399Arg was 1.48 (95% CI 5 1.06-2.07) using 194Arg-280Arg-399Arg as the reference. The OR for the 194Arg-280His-399Arg and the 194Arg-280Arg-399Gln were 1.78 (95% CI 5 1.09-2.89) and 1.78 (95% CI 5 1.23-2.59), respectively. Analysis after controlling for smoking, exercise and dietary habits indicated that alcohol consumption ( 80 g/week) is a significant risk factor of colorectal cancer (OR 5 2.60, 95% CI 5 1. 46-4.62). An increased risk for colorectal cancer was identified in alcohol drinkers with the risky allele combinations. Our results suggest that polymorphisms in the XRCC1 genes may contribute to colorectal cancer susceptibility, and some evidence was obtained of a genetic modification for the relationship between alcohol intake and colorectal cancer. ' 2005 Wiley-Liss, Inc.Key words: polymorphisms; XRCC1; genotype; allele; colorectal cancer; alcohol Colorectal cancer is one of the most commonly diagnosed cancers in North America and Western Europe, 1 and is the fourth most common cause of cancer in South Korea. 2 Inherited deficiencies in DNA repair have been associated with an individual's susceptibility to cancer. 3 Therefore, polymorphisms of DNA repair genes may increase the risk of colorectal cancer.The human XRCC1 gene, one of the DNA repair genes, was identified because of its ability to restore DNA repair activity in a Chinese hamster ovary mutant cell line EM9. 4 The XRCC1 protein is involved in base-excision repair, and interacts with DNA ligase III, DNA polymerase b, poly(ADP-ribose)polymerase (PARP), polynucleotide kinase and AP endonuclease I. 5-8 The base-excision repair pathway is designed to remove non-bulky base adducts, which are produced by methylation, oxidation or reduction by ionizing radiation or oxidative damage. 9,10 Three coding polymorphisms of the DNA repair gene XRCC1 (Arg194Trp, Arg280His and Arg399Gln) have been identified in man, suggesting altered efficiency due to amino acid substitutions. 11,12 Alcohol consumption has been associated with the production of reactive oxygen species, which are known to cause DNA lesions that can be removed by the DNA base-excision repair pathway. 13 Meta-analyses of alcohol consumption in relation to colorectal cancer have reported a small or moderate addit...

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“…23 Shen et al 11 reported 3 coding polymorphisms, i.e., Arg194Trp, Arg280His and Arg399Gln, in the XRCC1 gene.…”

Section: Discussionmentioning

confidence: 99%

Polymorphisms of XRCC1 gene, alcohol consumption and colorectal cancer

Hong

Lee

Kim

et al. 2005

Intl Journal of Cancer

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“…Soon thereafter, a stable complex of XRCC1 and LigIIIα was discovered [65] and by the late 1990s XRCC1 was established as a BER scaffold protein [66] in partnership with pol ß and PARP1 [16]. The list of cellular components that interact with XRCC1 has been expanded to include nicked DNA [67,68] and AP site containing DNA, both intermediates in the BER pathway [69].…”

Section: Scaffold Proteinsmentioning

confidence: 99%

“…Table I Possible entry mechanisms of a unified base excision repair model [12,17,23,63,66,75,88,113,119,128,129,149,150,[152][153][154][155][156][157] LigI…”

Section: Fig 2 a Unified Model Of Bermentioning

confidence: 99%

A unified view of base excision repair: Lesion-dependent protein complexes regulated by post-translational modification

Almeida¹,

Sobol²

2007

DNA Repair

385

374

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Base excision repair (BER) proteins act upon a significantly broad spectrum of DNA lesions that result from endogenous and exogenous sources. Multiple sub-pathways of BER (short-path or longpatch) and newly designated DNA repair pathways (e.g., SSBR and NIR) that utilize BER proteins complicate any comprehensive understanding of BER and its role in genome maintenance, chemotherapeutic response, neurodegeneration, cancer or aging. Herein, we propose a unified model of BER, comprised of three functional processes: Lesion Recognition/Strand Scission, Gap Tailoring and DNA Synthesis/Ligation, each represented by one or more multiprotein complexes and coordinated via the XRCC1/DNA Ligase III and PARP1 scaffold proteins. BER therefore may be represented by a series of repair complexes that assemble at the site of the DNA lesion and mediates repair in a coordinated fashion involving protein-protein interactions that dictate subsequent steps or sub-pathway choice. Complex formation is influenced by post-translational protein modifications that arise from the cellular state or the DNA damage response, providing an increase in specificity and efficiency to the BER pathway. In this review, we have summarized the reported BER proteinprotein interactions and protein post-translational modifications and discuss the impact on DNA repair capacity and complex formation.

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“…Studies continued to identify a role for pol β in the repair of damage induced by many different DNA damaging agents, including bleomycin [14,15] [21][22][23][24]. Several groups reported complete BER in vitro with pol β and additional purified proteins [22,23,25]. Although it was demonstrated in heterologous systems (E. coli and Saccharomyces cerevisiae) that pol β can conduct DNA replication and repair in vivo [26,27], it was not until a mouse gene knockout was made that the specificity of the repair conducted by pol β was defined [28].…”

Section: Dear Editormentioning

confidence: 99%

DNA polymerase beta null mouse embryonic fibroblasts harbor a homozygous null mutation in DNA polymerase iota

Sobol

2007

DNA Repair

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Dear EditorPrior to the explosion of gene targeting technology, considerable biochemical and cell-based experiments suggested a role for DNA polymerase β (pol β) in DNA repair. Since its initial discovery in 1971 [1], the enzyme that was to eventually be designated pol β [2,3] was unique in its enzymatic properties [4][5][6] as compared to the other newly characterized mammalian DNA polymerases alpha (pol α), gamma (pol γ) and delta (polδ) [1,7]. Of the four eukaryotic DNA polymerases identified by 1977, pol β soon earned the moniker of 'the' DNA repair polymerase [8][9][10][11]. These early studies defined a role for pol β in repair using isolated nuclei or nuclear extracts, monitoring the incorporation of radioactive nucleosides following ultraviolet (UV) light-induced DNA damage [8][9][10][11]. A more definitive role for pol β in excision repair in vitro was demonstrated in 1983, in which complete removal of pyrimidine dimers in DNA was conducted by purified enzyme preparations of pol β, T4 denV (UV endonuclease) and HeLa AP endonuclease II [12]. Although it was subsequently shown that pol α can also carry-out gap-filling DNA synthesis in a base excision repair (BER) reaction [13], the evidence continued to mount in support of pol β acting as 'the' DNA repair polymerase in the nucleus, while pol γ participated in replication and repair in mitochondria [1]. Studies continued to identify a role for pol β in the repair of damage induced by many different DNA damaging agents, including bleomycin [14,15] [21][22][23][24]. Several groups reported complete BER in vitro with pol β and additional purified proteins [22,23,25]. Although it was demonstrated in heterologous systems (E. coli and Saccharomyces cerevisiae) that pol β can conduct DNA replication and repair in vivo [26,27], it was not until a mouse gene knockout was made that the specificity of the repair conducted by pol β was defined [28].Characterization of the pol β knockout mouse [29,30] [34,54,55]; among other studies too numerous to mention herein. The most definitive and reproducible endpoint that has been used to evaluate pol β *To whom correspondence should be addressed: Robert W. Sobol, Hillman Cancer Center, University of Pittsburgh Cancer Institute, Research Pavilion, Suite 2.6, 5117 Centre Avenue, Pittsburgh, PA 15213-1863, Phone: 412-623-7764, Fax: 412-623-7761, e-mail: rws9@pitt.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript participation in repair in vivo is survival following DNA damage such as exposure to alkylating agen...

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Reconstitution of DNA base excision-repair with purified human proteins: interaction between DNA polymerase beta and the XRCC1 protein.

Cited by 707 publications

References 40 publications

Polymorphisms of XRCC1 gene, alcohol consumption and colorectal cancer

Polymorphisms of XRCC1 gene, alcohol consumption and colorectal cancer

A unified view of base excision repair: Lesion-dependent protein complexes regulated by post-translational modification

DNA polymerase beta null mouse embryonic fibroblasts harbor a homozygous null mutation in DNA polymerase iota

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