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Uracil dna glycosylase

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https://www.readbyqxmd.com/read/27875297/glycogen-synthase-kinase-3-gsk-3-mediated-phosphorylation-of-uracil-n-glycosylase-2-ung2-facilitates-repair-of-floxuridine-induced-dna-lesions-and-promotes-cell-survival
#1
Carly A Baehr, Catherine J Huntoon, Song-My Hoang, Calvin R Jerde, Larry M Karnitz
Uracil N-glycosylase 2 (UNG2), the nuclear isoform of UNG, catalyzes the removal of uracil or 5-fluorouracil lesions that accumulate in DNA following treatment with the anti-cancer agents 5-fluorouracil and 5-fluorodeoxyuridine (floxuridine), a 5-fluorouracil metabolite. By repairing these DNA lesions before they can cause cell death, UNG2 promotes cancer cell survival and is therefore critically involved in tumor resistance to these agents. However, the mechanisms by which UNG2 is regulated remain unclear...
November 14, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27871818/oxidatively-generated-base-modifications-in-dna-not-only-carcinogenic-risk-factor-but-also-regulatory-mark
#2
Marco Seifermann, Bernd Epe
The generation of DNA modifications in cells is in most cases accidental and associated with detrimental consequences such as increased mutation rates and an elevated risk of malignant transformation. Accordingly, repair enzymes involved in the removal of the modifications have primarily a protective function. Among the well-established exceptions of this rule are 5-methylcytosine and uracil, which are generated in DNA enzymatically under controlled conditions and fulfill important regulatory functions in DNA as epigenetic marks and in antibody diversification, respectively...
November 18, 2016: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/27836975/a-conserved-tripeptide-sequence-at-the-c-terminus-of-the-poxvirus-dna-processivity-factor-d4-is-essential-for-protein-integrity-and-function
#3
Manunya Nuth, Hancheng Guan, Robert P Ricciardi
Vaccinia virus (VACV) is a poxvirus member, and the VACV D4 protein serves both as a uracil-DNA glycosylase (UDG) and as an essential component required for processive DNA synthesis. The VACV A20 protein has no known catalytic function itself, but associates with D4 to form the D4-A20 heterodimer that functions as the poxvirus DNA processivity factor. The heterodimer enables the DNA polymerase to efficiently synthesize extended strands of DNA. Upon characterizing the interaction between D4 and A20, we observed that the C-terminus of D4 is susceptible to perturbation...
November 11, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27825529/toehold-mediated-strand-displacement-reaction-dependent-fluorescent-strategy-for-sensitive-detection-of-uracil-dna-glycosylase-activity
#4
Yushu Wu, Lei Wang, Wei Jiang
Sensitive detection of uracil-DNA glycosylase (UDG) activity is beneficial for evaluating the repairing process of DNA lesions. Here, toehold-mediated strand displacement reaction (TSDR)-dependent fluorescent strategy was constructed for sensitive detection of UDG activity. A single-stranded DNA (ssDNA) probe with two uracil bases and a trigger sequence were designed. A hairpin probe with toehold domain was designed, and a reporter probe was also designed. Under the action of UDG, two uracil bases were removed from ssDNA probe, generating apurinic/apyrimidinic (AP) sites...
October 21, 2016: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27815903/analysis-of-nuclear-uracil-dna-glycosylase-nudg-turnover-during-the-cell-cycle
#5
Jennifer A Fischer, Salvatore J Caradonna
Uracil-DNA glycosylases (UDG/UNG) are enzymes that remove uracil from DNA and initiate base-excision repair. These enzymes play a key role in maintaining genomic integrity by reducing the mutagenic events caused by G:C to A:T transition mutations. The recent finding that a family of RNA editing enzymes (AID/APOBECs) can deaminate cytosine in DNA has raised the interest in these base-excision repair enzymes. The methodology presented here focuses on determining the regulation of the nuclear isoform of uracil-DNA glycosylase (nUDG), a 36,000 Da protein...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27723754/targeted-aid-mediated-mutagenesis-tam-enables-efficient-genomic-diversification-in-mammalian-cells
#6
Yunqing Ma, Jiayuan Zhang, Weijie Yin, Zhenchao Zhang, Yan Song, Xing Chang
A large number of genetic variants have been associated with human diseases. However, the lack of a genetic diversification approach has impeded our ability to interrogate functions of genetic variants in mammalian cells. Current screening methods can only be used to disrupt a gene or alter its expression. Here we report the fusion of activation-induced cytidine deaminase (AID) with nuclease-inactive clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (dCas9) for efficient genetic diversification, which enabled high-throughput screening of functional variants...
October 10, 2016: Nature Methods
https://www.readbyqxmd.com/read/27721668/archaeal-dna-polymerase-b-as-a-dna-template-guardian-links-between-polymerases-and-base-alternative-excision-repair-enzymes-in-handling-the-deaminated-bases-uracil-and-hypoxanthine
#7
Javier Abellón-Ruiz, Sonoko Ishino, Yoshizumi Ishino, Bernard A Connolly
In Archaea repair of uracil and hypoxanthine, which arise by deamination of cytosine and adenine, respectively, is initiated by three enzymes: Uracil-DNA-glycosylase (UDG, which recognises uracil); Endonuclease V (EndoV, which recognises hypoxanthine); and Endonuclease Q (EndoQ), (which recognises both uracil and hypoxanthine). Two archaeal DNA polymerases, Pol-B and Pol-D, are inhibited by deaminated bases in template strands, a feature unique to this domain. Thus the three repair enzymes and the two polymerases show overlapping specificity for uracil and hypoxanthine...
2016: Archaea: An International Microbiological Journal
https://www.readbyqxmd.com/read/27704075/a-dna-array-based-on-clickable-lesion-containing-hairpin-probes-for-multiplexed-detection-of-base-excision-repair-activities
#8
Mélanie Flaender, Guillaume Costa, Guillaume Nonglaton, Christine Saint-Pierre, Didier Gasparutto
DNA is under continuous assault by environmental and endogenous reactive oxygen and alkylating species, inducing the formation of mutagenic, toxic and genome destabilizing nucleobase lesions. Due to the implications of such genetic alterations in cell death, aging, inflammation, neurodegenerative diseases and cancer, many efforts have been devoted to developing assays that aim at analyzing DNA repair activities from purified enzymes or cell extracts. The present work deals with the conception and application of a new, miniaturized and parallelized on surface-DNA biosensor to measure base excision repair (BER) activities...
September 22, 2016: Analyst
https://www.readbyqxmd.com/read/27697833/ung-protects-b-cells-from-aid-induced-telomere-loss
#9
Elena M Cortizas, Astrid Zahn, Shiva Safavi, Joseph A Reed, Francisco Vega, Javier M Di Noia, Ramiro E Verdun
Activation-induced deaminase (AID) initiates antibody gene diversification by creating G:U mismatches in the immunoglobulin loci. However, AID also deaminates nonimmunoglobulin genes, and failure to faithfully repair these off-target lesions can cause B cell lymphoma. In this study, we identify a mechanism by which processing of G:U produced by AID at the telomeres can eliminate B cells at risk of genomic instability. We show that telomeres are off-target substrates of AID and that B cell proliferation depends on protective repair by uracil-DNA glycosylase (UNG)...
October 17, 2016: Journal of Experimental Medicine
https://www.readbyqxmd.com/read/27684934/poxvirus-uracil-dna-glycosylase-an-unusual-member-of-the-family-i-uracil-dna-glycosylases
#10
REVIEW
Norbert Schormann, Natalia Zhukovskaya, Gregory Bedwell, Manunya Nuth, Richard Gillilan, Peter E Prevelige, Robert P Ricciardi, Surajit Banerjee, Debasish Chattopadhyay
Uracil-DNA glycosylases are ubiquitous enzymes, which play a key role repairing damages in DNA and in maintaining genomic integrity by catalyzing the first step in the base excision repair pathway. Within the superfamily of uracil-DNA glycosylases family I enzymes or UNGs are specific for recognizing and removing uracil from DNA. These enzymes feature conserved structural folds, active site residues and use common motifs for DNA binding, uracil recognition and catalysis. Within this family the enzymes of poxviruses are unique and most remarkable in terms of amino acid sequences, characteristic motifs and more importantly for their novel non-enzymatic function in DNA replication...
December 2016: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/27640531/actinomyces-spp-gene-expression-in-root-caries-lesions
#11
Naile Dame-Teixeira, Clarissa Cavalcanti Fatturi Parolo, Marisa Maltz, Aradhna Tugnait, Deirdre Devine, Thuy Do
BACKGROUND: The studies of the distribution of Actinomyces spp. on carious and non-carious root surfaces have not been able to confirm the association of these bacteria with root caries, although they were extensively implicated as a prime suspect in root caries. OBJECTIVE: The aim of this study was to observe the gene expression of Actinomyces spp. in the microbiota of root surfaces with and without caries. DESIGN: The oral biofilms from exposed sound root surface (SRS; n=10) and active root caries (RC; n=30) samples were collected...
2016: Journal of Oral Microbiology
https://www.readbyqxmd.com/read/27599859/structural-analysis-of-point-mutations-at-the-vaccinia-virus-a20-d4-interface
#12
Céline Contesto-Richefeu, Nicolas Tarbouriech, Xavier Brazzolotto, Wim P Burmeister, Christophe N Peyrefitte, Frédéric Iseni
The Vaccinia virus polymerase holoenzyme is composed of three subunits: E9, the catalytic DNA polymerase subunit; D4, a uracil-DNA glycosylase; and A20, a protein with no known enzymatic activity. The D4/A20 heterodimer is the DNA polymerase cofactor, the function of which is essential for processive DNA synthesis. The recent crystal structure of D4 bound to the first 50 amino acids of A20 (D4/A201-50) revealed the importance of three residues, forming a cation-π interaction at the dimerization interface, for complex formation...
September 2016: Acta Crystallographica. Section F, Structural Biology Communications
https://www.readbyqxmd.com/read/27591637/label-free-fluorescence-turn-on-detection-of-uracil-dna-glycosylase-activity-based-on-g-quadruplex-formation
#13
Changbei Ma, Kefeng Wu, Haisheng Liu, Kun Xia, Kemin Wang, Jun Wang
We have developed a new methodology for fluorescence turn-on detection of uracil DNA glycosylase (UDG) activity based on G-quadruplex formation using a thioflavin T probe. In the presence of UDG, it catalyzed the hydrolysis of the uracil bases in the duplex DNA, resulting in the dissociation of the duplex DNA owing to their low melting temperature. Then, the probe DNA can be recognized quickly by the ThT dye and resulting in an increase in fluorescence. This approach is highly selective and sensitive with a detection limit of 0...
November 1, 2016: Talanta
https://www.readbyqxmd.com/read/27580719/structural-basis-of-damage-recognition-by-thymine-dna-glycosylase-key-roles-for-n-terminal-residues
#14
Christopher T Coey, Shuja S Malik, Lakshmi S Pidugu, Kristen M Varney, Edwin Pozharski, Alexander C Drohat
Thymine DNA Glycosylase (TDG) is a base excision repair enzyme functioning in DNA repair and epigenetic regulation. TDG removes thymine from mutagenic G·T mispairs arising from deamination of 5-methylcytosine (mC), and it processes other deamination-derived lesions including uracil (U). Essential for DNA demethylation, TDG excises 5-formylcytosine and 5-carboxylcytosine, derivatives of mC generated by Tet (ten-eleven translocation) enzymes. Here, we report structural and functional studies of TDG(82-308), a new construct containing 29 more N-terminal residues than TDG(111-308), the construct used for previous structures of DNA-bound TDG...
December 1, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27571472/comparative-effects-of-ions-molecular-crowding-and-bulk-dna-on-the-damage-search-mechanisms-of-hogg1-and-hung
#15
Shannen L Cravens, James T Stivers
The energetic nature of the interactions of DNA base excision repair glycosylases with undamaged and damaged DNA and the nuclear environment are expected to significantly impact the time it takes for these enzymes to search for damaged DNA bases. In particular, the high concentration of monovalent ions, macromolecule crowding, and densely packed DNA chains in the cell nucleus could alter the search mechanisms of these enzymes as compared to findings in dilute buffers typically used in in vitro experiments. Here we utilize an in vitro system where the concerted effects of monovalent ions, macromolecular crowding, and high concentrations of bulk DNA chains on the activity of two paradigm human DNA glycosylases can be determined...
September 20, 2016: Biochemistry
https://www.readbyqxmd.com/read/27571178/the-ddb1-dcaf1-vpr-ung2-crystal-structure-reveals-how-hiv-1-vpr-steers-human-ung2-toward-destruction
#16
Ying Wu, Xiaohong Zhou, Christopher O Barnes, Maria DeLucia, Aina E Cohen, Angela M Gronenborn, Jinwoo Ahn, Guillermo Calero
The HIV-1 accessory protein Vpr is required for efficient viral infection of macrophages and promotion of viral replication in T cells. Vpr's biological activities are closely linked to the interaction with human DCAF1, a cellular substrate receptor of the Cullin4-RING E3 ubiquitin ligase (CRL4) of the host ubiquitin-proteasome-mediated protein degradation pathway. The molecular details of how Vpr usurps the protein degradation pathway have not been delineated. Here we present the crystal structure of the DDB1-DCAF1-HIV-1-Vpr-uracil-DNA glycosylase (UNG2) complex...
October 2016: Nature Structural & Molecular Biology
https://www.readbyqxmd.com/read/27517750/inhibition-of-uracil-dna-glycosylase-sensitizes-cancer-cells-to-5-fluorodeoxyuridine-through-replication-fork-collapse-induced-dna-damage
#17
Yan Yan, Xiangzi Han, Yulan Qing, Allison G Condie, Shashank Gorityala, Shuming Yang, Yan Xu, Youwei Zhang, Stanton L Gerson
5-fluorodeoxyuridine (5-FdU, floxuridine) is active against multiple cancers through the inhibition of thymidylate synthase, which consequently introduces uracil and 5-FU incorporation into the genome. Uracil DNA glycosylase (UDG) is one of the main enzymes responsible for the removal of uracil and 5-FU. However, how exactly UDG mediates cellular sensitivity to 5-FdU, and if so whether it is through its ability to remove uracil and 5-FU have not been well characterized. In this study, we report that UDG depletion led to incorporation of uracil and 5-FU in DNA following 5-FdU treatment and significantly enhanced 5-FdU's cytotoxicity in cancer cell lines...
August 9, 2016: Oncotarget
https://www.readbyqxmd.com/read/27499380/aptamer-mediated-universal-enzyme-assay-based-on-target-triggered-dna-polymerase-activity
#18
Ki Soo Park, Chang Yeol Lee, Kyoung Suk Kang, Hyun Gyu Park
We herein describe an innovative method for a universal fluorescence turn-on enzyme assay, which relies on the target enzyme-triggered DNA polymerase activity. In the first target recognition step, the target enzyme is designed to destabilize detection probe derived from an aptamer specific to DNA polymerase containing the overhang sequence and the complementary blocker DNA, which consequently leads to the recovery of DNA polymerase activity inhibited by the detection probe. This target-triggered polymerase activity is monitored in the second signal transduction step based on primer extension reaction coupled with TaqMan probe...
July 12, 2016: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27492474/targeted-nucleotide-editing-using-hybrid-prokaryotic-and-vertebrate-adaptive-immune-systems
#19
Keiji Nishida, Takayuki Arazoe, Nozomu Yachie, Satomi Banno, Mika Kakimoto, Mayura Tabata, Masao Mochizuki, Aya Miyabe, Michihiro Araki, Kiyotaka Y Hara, Zenpei Shimatani, Akihiko Kondo
The generation of genetic variation (somatic hypermutation) is an essential process for the adaptive immune system in vertebrates. We demonstrate the targeted single-nucleotide substitution of DNA using hybrid vertebrate and bacterial immune systems components. Nuclease-deficient type II CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated) and the activation-induced cytidine deaminase (AID) ortholog PmCDA1 were engineered to form a synthetic complex (Target-AID) that performs highly efficient target-specific mutagenesis...
September 16, 2016: Science
https://www.readbyqxmd.com/read/27479048/label-free-thioflavin-t-g-quadruplex-based-real-time-strand-displacement-amplification-for-biosensing-applications
#20
Yi-Chen Du, Li-Na Zhu, De-Ming Kong
To promote application of strand-displacement amplification (SDA) techniques in biosensing, a label-free, real-time monitoring strategy for isothermal nucleic acid amplification reactions was designed. G-quadruplex structures were introduced into SDA products using specific recognition of G-quadruplexes by the fluorogenic dye thioflavin T. Performance was good for real-time monitoring of traditional SDA by a linear-amplification mechanism and for exponential cross-triggered SDA amplification. The strategy worked on a commercial real-time PCR instrument, making it suitable for biosensing platforms...
December 15, 2016: Biosensors & Bioelectronics
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