Determination of Glycated Nucleobases in Human Urine by a New Monoclonal Antibody Specific for <i>N</i><sup>2</sup>-Carboxyethyl-2‘-deoxyguanosine

Schneider, M. U.; Thoss, Gerlinde; Hübner-Parajsz, Christa; Kientsch-Engel, Rosemarie; Stahl, Peter J.; Pischetsrieder, Monika

doi:10.1021/tx049929d

Cited by 39 publications

(41 citation statements)

References 25 publications

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“…caused by endogenous agents that similarly require Rev1's polymerase activity for bypass. An example of such an endogenous agent produced by normal metabolism is methylglyoxal, a reduced form of pyruvic acid that can react with DNA to produce N 2 -(1-carboxyethyl)-29-deoxyguanosine (N 2 -CEdG) as the major lesion (Frischmann et al 2005;Yuan et al 2008), which is also detectable in human samples (Schneider et al 2004;Li et al 2006). We assessed the percentage of survival of our strains in the presence of chronic exposure to methylglyoxal, choosing the rev1Dmms2D strain background for our experiments to maximize any observable phenotypes for the catalytic dead rev1 mutant.…”

Section: Resultsmentioning

confidence: 99%

The DNA Polymerase Activity ofSaccharomyces cerevisiaeRev1 is Biologically Significant

Wiltrout¹,

Walker

2011

Genetics

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A cell's ability to tolerate DNA damage is directly connected to the human development of diseases and cancer. To better understand the processes underlying mutagenesis, we studied the cell's reliance on the potentially error-prone translesion synthesis (TLS), and an error-free, template-switching pathway in Saccharomyces cerevisiae. The primary proteins mediating S. cerevisiae TLS are three DNA polymerases (Pols): Rev1, Pol z (Rev3/7), and Pol h (Rad30), all with human homologs. Rev1's noncatalytic role in recruiting other DNA polymerases is known to be important for TLS. However, the biological significance of Rev1's unusual conserved DNA polymerase activity, which inserts dC, is much less well understood. Here, we demonstrate that inactivating Rev1's DNA polymerase function sensitizes cells to both chronic and acute exposure to 4-nitroquinoline-1-oxide (4-NQO) but not to UV or cisplatin. Full Rev1-dependent resistance to 4-NQO, however, also requires the additional Rev1 functions. When error-free tolerance is disrupted through deletion of MMS2, Rev1's catalytic activity is more vital for 4-NQO resistance, possibly explaining why the biological significance of Rev1's catalytic activity has been elusive. In the presence or absence of Mms2-dependent error-free tolerance, the catalytic dead strain of Rev1 exhibits a lower 4-NQO-induced mutation frequency than wild type. Furthermore, Pol z, but not Pol h, also contributes to 4-NQO resistance. These results show that Rev1's catalytic activity is important in vivo when the cell has to cope with specific DNA lesions, such as N 2 -dG.

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Section: Resultsmentioning

confidence: 99%

The DNA Polymerase Activity ofSaccharomyces cerevisiaeRev1 is Biologically Significant

Wiltrout¹,

Walker

2011

Genetics

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“…It was found recently that MG induces modifications in calf thymus DNA mainly on dG to give N 2 -CEdG (22). N 2 -CEdG could also be detected in urine samples of healthy human subjects (23) and in kidney and aorta cells of diabetic and uremic patients (24). LC-MS/MS with the isotope dilution method revealed that N 2 -CEdG can be formed in untreated WM-266-4 cells at a level of approximately one lesion per 10 7 nucleosides; treatment of cells with MG or glucose can further enhance the formation of the lesion, supporting that N 2 -CEdG is an endogenous DNA lesion, and the amount of the lesion can be increased by byproducts of glycolysis.…”

Section: Discussionmentioning

confidence: 98%

“…1) (22). A competitive enzyme-linked immunosorbant assay showed that the adduct can be detected in urine samples of 121 healthy human subjects at levels ranging from 1.2-to 117-ng N 2 -CEdG equivalent per milligram of creatinine (23). Immunohistochemistry using a monoclonal antibody against N 2 -CEdG revealed that the level of the lesion is enhanced in the kidney and aorta of patients with diabetic nephropathy and uremic atherosclerosis, respectively (24).…”

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confidence: 99%

Efficient and accurate bypass of N ² -(1-carboxyethyl)-2′-deoxyguanosine by DinB DNA polymerase in vitro and in vivo

Yuan¹,

Cao²,

Jiang

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

135

153

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DinB, a Y-family DNA polymerase, is conserved among all domains of life; however, its endogenous substrates have not been identified. DinB is known to synthesize accurately across a number of N 2 -dG lesions. Methylglyoxal (MG) is a common byproduct of the ubiquitous glycolysis pathway and induces the formation of N 2 -(1-carboxyethyl)-2-deoxyguanosine (N 2 -CEdG) as the major stable DNA adduct. Here, we found that N 2 -CEdG could be detected at a frequency of one lesion per 10 7 nucleosides in WM-266-4 human melanoma cells, and treatment of these cells with MG or glucose led to a dose-responsive increase in N 2 -CEdG formation. We further constructed single-stranded M13 shuttle vectors harboring individual diastereomers of N 2 -CEdG at a specific site and assessed the cytotoxic and mutagenic properties of the lesion in wild-type and bypass polymerase-deficient Escherichia coli cells. Our results revealed that N 2 -CEdG is weakly mutagenic, and DinB (i.e., polymerase IV) is the major DNA polymerase responsible for bypassing the lesion in vivo. Moreover, steady-state kinetic measurements showed that nucleotide insertion, catalyzed by E. coli pol IV or its human counterpart (i.e., polymerase ), opposite the N 2 -CEdG is both accurate and efficient. Taken together, our data support that N 2 -CEdG, a minor-groove DNA adduct arising from MG, is an important endogenous substrate for DinB DNA polymerase.glycolysis ͉ mutagenesis ͉ polymerase ͉ translesion synthesis

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“…1) (16). This modified nucleoside was found to be present in urine samples from healthy human subjects (17), and the lesion was observed more frequently in kidney and aortic cells of diabetic and uremic patients than in the corresponding cells of healthy human subjects (18). N 2 -CEdG can also be detected in cultured human cells, and the exposure of these cells to methylglyoxal or glucose further enhanced the formation of this lesion (14).…”

mentioning

confidence: 96%

The Roles of DNA Polymerases κ and ι in the Error-free Bypass of N2-Carboxyalkyl-2′-deoxyguanosine Lesions in Mammalian Cells

Yuan

You²,

Andersen³

et al. 2011

Journal of Biological Chemistry

101

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To counteract the deleterious effects of DNA damage, cells are equipped with specialized polymerases to bypass DNA lesions. Previous biochemical studies revealed that DinB family DNA polymerases, including Escherichia coli DNA polymerase IV and human DNA polymerase , efficiently incorporate the correct nucleotide opposite some N DNA is susceptible to damage by endogenous and exogenous agents (1). To minimize cell death arising from replication blockage and mutations emanating from nucleotide misincorporation opposite the damage site, cells are equipped with several pathways to repair DNA lesions (2). Additionally, cells have evolved two major mechanisms to tolerate unrepaired DNA lesions (3), including homologous recombination and translesion synthesis (TLS) 2 (3). In this regard, synthesis past many DNA lesions requires the replacement of replicative DNA polymerases with one or a few specialized polymerases, most of which belong to the Y-family (4). Along this line, phylogenetic analysis revealed five branches of Y-family polymerases, which include UmuC, DinB, polymerase , polymerase , and REV1 (4, 5). All of these, except for UmuC, are found in eukaryotic cells (4,5). Apart from the Y-family DNA polymerases, some B-family DNA polymerases also participate in TLS. These include DNA polymerase II in Escherichia coli and polymerase , which is composed of the catalytic subunit REV3 and the regulatory subunit REV7, in eukaryotic cells (4, 5). These specialized DNA polymerases are characterized by their relatively low fidelity in replicating undamaged DNA but have the capability to bypass those DNA lesions that normally block DNA synthesis by replicative DNA polymerases (4). It was advocated that specialized DNA polymerases are evolved by nature to copy DNA base damage accurately, but they no longer bear the ability to replicate unmodified DNA with high fidelity (6). The best known example of this is the efficient and accurate bypass of cis,syn-cyclobutane pyrimidine dimers by polymerase (7,8), which is encoded by the xeroderma pigmentosum-variant gene (i.e. POLH) in humans (9). Xeroderma pigmentosum-variant patients exhibit elevated susceptibility to developing skin cancer (10). Several recent studies also demonstrated that DinB DNA polymerase, a Y-family polymerase conserved in all three kingdoms of life (11), can insert the correct dCMP opposite several N 2 -substituted guanine lesions at an efficiency that is similar to or better than opposite an unmodified guanine (12)(13)(14). The x-ray crystal structure of the catalytic core of human DNA polymerase , along with primer/template DNA and an incoming nucleotide, reveals the lack of steric hindrance in the minor groove at the primer-template junction (15), which may account for the tolerance of polymerase toward the minor groove N 2 -dG lesions. By using shuttle vector technology, we also showed that DinB (i.e. polymerase IV) is the major polymerase involved in the accurate bypass of the two diastereomers of N 2 -(1-carboxyethyl)-2Ј-deoxyguanosine (N 2 -CEdG) in ...

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Determination of Glycated Nucleobases in Human Urine by a New Monoclonal Antibody Specific for N²-Carboxyethyl-2‘-deoxyguanosine

Cited by 39 publications

References 25 publications

The DNA Polymerase Activity ofSaccharomyces cerevisiaeRev1 is Biologically Significant

The DNA Polymerase Activity ofSaccharomyces cerevisiaeRev1 is Biologically Significant

Efficient and accurate bypass of N ² -(1-carboxyethyl)-2′-deoxyguanosine by DinB DNA polymerase in vitro and in vivo

The Roles of DNA Polymerases κ and ι in the Error-free Bypass of N2-Carboxyalkyl-2′-deoxyguanosine Lesions in Mammalian Cells

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Determination of Glycated Nucleobases in Human Urine by a New Monoclonal Antibody Specific for N2-Carboxyethyl-2‘-deoxyguanosine

Cited by 39 publications

References 25 publications

The DNA Polymerase Activity ofSaccharomyces cerevisiaeRev1 is Biologically Significant

The DNA Polymerase Activity ofSaccharomyces cerevisiaeRev1 is Biologically Significant

Efficient and accurate bypass of N 2 -(1-carboxyethyl)-2′-deoxyguanosine by DinB DNA polymerase in vitro and in vivo

The Roles of DNA Polymerases κ and ι in the Error-free Bypass of N2-Carboxyalkyl-2′-deoxyguanosine Lesions in Mammalian Cells

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Product

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Determination of Glycated Nucleobases in Human Urine by a New Monoclonal Antibody Specific for N²-Carboxyethyl-2‘-deoxyguanosine

Efficient and accurate bypass of N ² -(1-carboxyethyl)-2′-deoxyguanosine by DinB DNA polymerase in vitro and in vivo