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4.2.99.18: DNA-(apurinic or apyrimidinic site) lyase

This is an abbreviated version!
For detailed information about DNA-(apurinic or apyrimidinic site) lyase, go to the full flat file.

Word Map on EC 4.2.99.18

Reaction

the C-O-P bond 3' to the apurinic or apyrimidinic site in DNA is broken by a beta-elimination reaction, leaving a 3'-terminal unsaturated sugar and a product with a terminal 5'-phosphate =

Synonyms

8-oxoG DNA glycosylase, abasic (AP)-endonuclease, abasic endonuclease, AdAPE1/Ref-1, Af0371, Afogg enzyme, ALKBH1, AP 1, AP DNA endonuclease 1, AP Dnase, AP endo, AP endonuclease, AP endonuclease 1, AP endonuclease Ape1, AP endonuclease Class I, AP endonuclease I, AP endonuclease VI, AP lyase, AP site-DNA 5'-phosphomonoester-lyase, AP-endonuclease, AP-endonuclease 1, AP-endonuclease 2, AP-endonuclease/3'-5'exodeoxyribonuclease, AP-lyase, Ap1, Ap2, Ape, APE-1, APE/Ref-1, APE1, APE1/Ref-1, Ape1L, Ape2, APEN, APEX nuclease, APEX1, APLM, APN, APN-1, APN1, apurinic DNA endonuclease, apurinic endodeoxyribonuclease, apurinic endonuclease, apurinic endonuclease 1, apurinic-apyrimidinic DNA endonuclease, apurinic-apyrimidinic endodeoxyribonuclease, apurinic-apyrimidinic endonuclease, apurinic-apyrimidinic endonuclease 1, apurinic-apyrimidinic endonuclease I, apurinic/apyrimidic endonuclease 1, apurinic/Apyrimidinic (AP) endonuclease 1, apurinic/apyrimidinic AP endonuclease, apurinic/apyrimidinic DNA endonuclease 1, apurinic/apyrimidinic endonuclease, apurinic/apyrimidinic endonuclease 1, apurinic/apyrimidinic endonuclease 1/redox factor-1, apurinic/apyrimidinic endonuclease 2, apurinic/apyrimidinic endonuclease APE1, apurinic/apyrimidinic endonuclease-1, apurinic/apyrimidinic endonuclease-1/Redox factor-1, apurinic/apyrimidinic endonuclease/redox effector factor-1, Apurinic/apyrimidinic endonuclease1/redox factor 1, apurinic/apyrimidinic endonuclease1/redox factor-1, apurinic/apyrimidinic lyase, apurinic/apyrimidinic site lyase, apurinic/apyrimidinic specific endonuclease, apurinic/apyrimidinic-endonuclease, apyrimidinic endonuclease, archaeal GO glycosylase, BAP1, class II AP endonuclease LMAP, class II apurinic/apyrimidinic(AP)-endonuclease, class-II AP-endonuclease, deoxyribonuclease (apurinic or apyrimidinic), DNA lyase, DNA polymerase X, DNA-(apurinic or apyrimidinic site) lyase, E. coli endonuclease III, EC 3.1.25.2, EcoNth, Endo III, endodeoxyribonuclease, endodeoxyribonuclease III, EndoIV, endonuclease III, endonuclease IV, endonuclease VI, endonuclease VIII, Escherichia coli endonuclease III, EXO-3, ExoA, ExoA type AP endonuclease, ExoIII, HAP-1, HAP1, HAP1h, hAPE, hAPE1, hNTH, hNTH1, hOgg1 protein, HpXth, human apurinic/apyrimidinic endonuclease, human apurinic/apyrimidinic endonuclease 1, Kae1, KsgA, LplExo, Micrococcus luteus UV endonuclease, MJ0724, mjOgg, MMH, More, mtAPE, MtbXthA, multi-functional apyrimidinic endonuclease1/redox factor-1, N-glycosylase AP lyase, N-glycosylase apurinic/apyrimidinic lyase, NapE, Nei, Nfo, Ntg1, Ntg1p, Ntg2, Ntg2p, Nth, NTH1, nuclease SmnA, nuclease, apurinic endodeoxyribo-, nuclease, apurinic-apyrimidinic endodeoxyribo-, nuclease, endodeoxyribo-, III, Ogg2, oxoG DNA glycosylase, Pa-AGOG, Pa-AGOG DNA glycosylase, PAE2237, PaKae1, PALF, pE296R, PF0258, phage-T4 UV endonuclease, redox effector factor 1, redox factor-1, redox factor-1:Ref-1, Ref-1, REF-1 protein, Ref1, SisExoIII, Smx nuclease, TvoExo, UL30, UV endo V, UV endonuclease, UV endonuclease V, X-ray endonuclease III, XTH, xthA, ZAP1, zApe

ECTree

     4 Lyases
         4.2 Carbon-oxygen lyases
             4.2.99 Other carbon-oxygen lyases
                4.2.99.18 DNA-(apurinic or apyrimidinic site) lyase

Cloned

Cloned on EC 4.2.99.18 - DNA-(apurinic or apyrimidinic site) lyase

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
adenoviruses full-length APE1/Ref-1 (AdAPE1/Ref-1) is generated by homologous recombination in human embryonic kidney 293 cells. Human umbilical vein endothelial cells are infected with 200 multiplicity of infection
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apn-1 gene of Caenorhabditis elegans expresses AP endonuclease activity in yeast mutants
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chromosomal DNA is isolated from Bacillus subtilis, and competent cells are prepared and transformed with plasmid or chromosomal DNA. nfo mutant strains are constructed
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DNA constructs encoding mutant Endo IV enzymes are generated with the megaprimer PCR technique47 using a wild-type nfo gene sequence as template. The PCR products are purified, digested and subcloned in pET24. Hexahistidine-tagged wild-type Endo IV and mutants are subcloned in vector pET28a. The double mutants (Y72A/E261Q and R37A/E261Q) are obtained by Megaprimer mutagenesis on the E261Q mutant followed by subcloning of the PCR product in frame with an N-terminal His6-sequence in pET28a. Wild-type and mutant enzymes are produced and purified using an nfo-negative strain BW565DE3 transformed with pET24-Endo IV constructs
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ectopical expression of FLAG-tagged wild-type APE1 and mutants K27Q or H309N in HEK-293T cell with downregulated endogenous APE1
exo-3 gene of Caenorhabditis elegans expresses AP endonuclease activity in yeast mutants
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expressed in Escherichia coli
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli Rosetta cells
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expressed in Escherichia coli Rosetta(DE3) cells
expressed in Escherichia coli strain BH110
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expression in Escherichia coli
expression in Sf9 cell
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expression vectors encoding the full-length zApe and zApe (33-310) proteins in pET15b are constructed using standard subcloning techniques and verified by DNA sequencing. The zApe vectors are transformed into Rosetta (DE3) Escherichia coli, cultures are grown in LB media
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for expression in Escherichia coli
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fusion protein with glutathione S-transferase
gene APE1, recombinant expression of enzyme in Escherichia coli strain Rosetta II(DE3)
gene APE1, recombinant expression of the enzyme in Escherichia coli strain Rosetta II(DE3)
gene APN1, sequence comparisons, recombinant expression of His-tagged wild-type and mutant enzymes
gene xth, DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant N-terminally His-tagged enzyme expression in Escherichia coli strains BH110 (DE3) and Rosetta 2 (DE3)
Generation of random mutations in the ape1 gene and selection of variants that confer protection against H2O2. Random library of ape1 mutants is generated by transforming the mutator strain XL1-Red with the prokaryotic ape1 plasmid pKK-ape1. The H2O2-resistant phenotype (mutant D70A) is expressed in Escherichia coli strain BW528 (xth nfo), which is deficient in the major AP endonucleases, exonuclease III and endonuclease IV
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His-tagged APE1 is expressed in Escherichia coli M15 cells
Human Ape1/Ref-1 cDNA is amplified by RT-PCR using Ape1/Ref-1-specific primers and from human GM00637 fibroblasts. AP endonuclease domain deletion mutant deltaAPD is amplified by RT-PCR using specific primers from full length human Ape1/Ref-1 cDNA. Redox domain deletion mutant deltaRD is amplified by RT-PCR using specific primers from full length human Ape1/Ref-1 cDNA.
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into the pET-14b vector
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into the pET15b vector for expression in Escherichia coli
into the pET15b vector for expression in Escherichia coli BL21DE3 cells
into the pET28a vector for expression in Escherichia coli BL21DE3 cells, into the pKK223-3 vector for transformation into the BW528 strain for subsequent complementation studies
into the pET28b+ vector for expression in Escherichia BL21DE3 RIL cells
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into the pProEX HTb vector for transformation of Escherichia coli XL-1 Blue and ER2566 cells
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into the pRSET-A vector for expression in Escherichia coli BL21DE3/pLys cells, the expression of pE296R protein is analyzed in ASFV-infected Vero cells
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into the pSLV vector to generate pSLX-APE-Myc
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into various expression plasmids for GFP, GST or His-tag, for expression in Escherichia coli cells
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into vector pET-24b, recombinant plasmids maintained in Escherichia coli XL1-Blue X cells, transfected into Escherichia L21(DE3)
mutant A138D is produced by site-directed mutagenesis and expressed in Escherichia coli strain BW528. Overexpression of LMAPA138D sensitizes Leishmania cells to oxidative agents.
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neuronal cell cultures are transfected with Api1 small interfering RNA, rat neuronal cell cultures are infected with adenovirus
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Ntg1p and Ntg2p
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overexpression in Escherichia coli
overproduced in Escherichia coli as a maltose binding protein(MBP)-Afogg fusion protein
recombinant enzyme expression in Escherichia coli strain Rosetta II(DE3)
recombinant expression of enzyme mutant Y105A
recombinant expression of wild-type and mutant enzymes
Recombinant human APE1 protein is overexpressed from clone pETApe1 in Escherichia coli BL21(DE3)
recombinant production of APE1
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repression of APE/Ref-1 by an antisense overexpression, transfection of mouse B-cells with phosphorothioate oligonucleotide, which is complementary to the APE/Ref-1 mRNA sequence and overlaps the translation initiation site (antisense oligonucleotide)
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RKO cell lines are established, that can be induced by doxycycline to overexpress APE-1 wild-type, mutant C65A, mutant E96A or mutant E96Q
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the APE-1/Ref-1 expression vector pFLAG-APE-1 cDNA3.1 is prepared
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The human NTH1 cDNA is cloned into the PCRII plasmid and it is cloned into the EcoRI site of the pGEX-2TK vector. The hNTH1 cDNA is cut (amino acids 1-94) and cloned into the modified restriction sites in the pGEX-2TK vector. The remaining fragment (amino acids 95-312) is cloned into the pGEX-3X vector. The hNTH1 cDNA is cloned into the pET24d vector in-frame with the His-tag and into the pNTAP-B vector in-frame with the TAP-epitope. To knock down the expression of Y-box-binding protein-1, a hairpin sequence specific to Y-box-binding protein-1 is cloned into the pSuper vector. His-hNTH1 protein is produced in the BL21 bacterial strain. MCF-7 cells are transfected with a siRNA specific to hNTH1.
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The pPS904 green fluorescent protein (GFP) expression vector (2m URA3) is employed for generation of C-terminally tagged Ntg1-GFP fusion protein. The Saccharomyces cerevisiae haploid strain FY86 is utilized for all localization studies. NTG1 strain (DSC0282) is generated by precisely replacing the NTG1 open reading frame in FY86 with the kanamycin antibiotic resistance gene. Haploid yeast strain expresses integrated genomic copies of C-terminally tandem affinity purification (TAP)-tagged Ntg1. Cells expressing galactose-inducible SMT3-HA and Ntg1-GST (DSC0221) are generated by integrating the hemagglutinin tag from the vector p1375 and the GAL promoter and glutathione S-transferase (GST) tag from the vector p2245 at the C-termini of the SMT3 and NTG1 products in the haploid strain ACY737.
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The pPS904 green fluorescent protein (GFP) expression vector (2m URA3) is employed for generation of C-terminally tagged Ntg2-GFP fusion protein. The Saccharomyces cerevisiae haploid strain FY86 is utilized for localization studies. NTG2 strain (DSC0283) is generated by precisely replacing NTG2 open reading frame in FY86 with the kanamycin antibiotic resistance gene. Haploid yeast strain expresses integrated genomic copies of C-terminally tandem affinity purification (TAP)-tagged Ntg2. Cells expressing galactose-inducible Smt3-HA and Ntg2-GST (DSC0222) are generated by integrating the hemagglutinin tag from the vector p1375 and the GAL promoter and glutathione S-transferase (GST) tag from the vector p2245 at the C-termini of the SMT3 and NTG2 products in the haploid strain ACY737.
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The wild-type and mutant hApe1 proteins are expressed as GST-fusions in Escherichia coli
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the yeast apurinic/apyrimidinic endonuclease gene is cloned into the pcDNA3.neo plasmid for transfection of rat RCSN-3 cells
UL30 is expressed in Spodoptera frugiperda cells
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wild-type APE1 and its Cys mutants are expressed as His-tag fusion polypeptides in Escherichia coli
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wild-type APE1 encoding full-length of APE1 and APE1 C65A/C93A encoding mutant form of APE1 in pCMVTag2B mammalian expression vector are generated by standard cloning method. TAT-APE1 is generated by insertion of full-length of APE1 into pTAT-2.1
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