Properties of DNA Repair Enzymes and Structure-specific Endonucleases

NEB carries an array of reliable DNA repair enzymes for use in multiple applications.

Enzyme Major Substrate1,2 Cleave site Product(s) Produced Termini Created from Cleavage Major Activity Thermostable
        5´ 3´    
APE 1  AP site 1st phosphodiester bond 5´ to AP site 1 nt gap dR5P  OH              Endonuclease  
Authenticase®
cruciform, mismatches, Holliday junctions, mismatches except G/A
phosphodiester bond 5´ and/or 3´ to structure


5´ and/or 3´ overhang
P OH
Endonuclease
 
Mismatch Endonuclease I
T:T, G:G and G:T mismatches in dsDNA
3rd phosphodiester bond on the 5´ side of the mismatched base in both strands 
5 bp overhang P  OH Endonuclease
 
T7 Endo I  crucifroms, mismatches, Holliday junctions, across DNA nicks phosphodiester bond 5´ to structure nick P              OH  Endonuclease  
Endo III  AP site, damaged pyrimidines, Tg N-glycosidic bond; 1st phosphodiester bond 3´ to AP site 1 nt gap P PA Glycosylase & AP lyase  
Tma Endo III AP site, damaged pyrimidines, Tg N-glycosidic bond; 1st phosphodiester bond 3´ to AP site  1 nt gap P PA Glycosylase & AP lyase YES
Endo IV AP site 1st phosphodiester bond 5´ to AP site 1 nt gap dR5P  OH              Endonuclease  
Tth Endo IV  AP site 1st phosphodiester bond 5´ to AP site  1 nt gap dR5P  OH              Endonuclease YES
Endo V  dI4, dU, AP site 2nd phosphodiester bond 3´ to dI nick P              OH              Endonuclease  
T4 PDG  CPD, AP site N-glycosidic bond; phosphodiester bond 3´ to AP site AP site; 1 nt gap P6 
Glycosylase & AP lyase               
Endo VIII  AP site4 phosphodiester bond 3´ & 5´ to AP site 1 nt gap P P AP lyase  
Thermostable EndoQ dU, dI, dX, AP site4 1st phosphodiester bond 5´ to modified nucleobase nick P              OH Endonuclease YES
Thermostable FEN1  5´ DNA flap3 phosphodiester bond at base of flap nick P              OH (on flap)  Endonuclease YES
Fpg  8-oxoG, oxidized purines N-glycosidic bond; phosphodiester bond 3´ & 5´ to AP site AP site; 1 nt gap P P Glycosylase & AP lyase  
hAAG  3mA, 7mG, dI, dX  N-glycosidic bond AP site N/A N/A Glycosylase  
hSMUG1  dU4, 5-hmU, 5-hoU, 5-fU N-glycosidic bond AP site N/A N/A Glycosylase  
Thermostable OGG  8oxoG  N-glycosidic bond; phosphodiester bond 3' & 5' to AP site  AP site; 1 nt gap  P  P  Glycosylase  YES 
RNaseHII              rN in dsDNA phosphodiester bond 5´ to ribo nick P              OH  Endonuclease  
UDG  dU4 N-glycosidic bond AP site N/A N/A Glycosylase  
Afu UDG  dU4 N-glycosidic bond AP site N/A N/A Glycosylase YES
Antarctic Thermolabile UDG5  dU4           N-glycosidic bond
AP site
N/A
N/A
Glycosylase  
WarmStart® Afu Uracil-DNA Glycosylase (UDG)
dU4 N-glycosidic bond AP site
N/A
N/A
Glycosylase
YES
USER Enzyme dU       N-glycosidic bond; phosphodiester bond 3´ & 5´ to AP site
1 nt gap
P  P
Glycosylase & AP lyase  
Thermolabile USER II dU         N-glycosidic bond; phosphodiester bond 3´ & 5´ to AP site
1 nt gap
P 
PA
Glycosylase & AP lyase  
Thermostable USER III dU       N-glycosidic bond; phosphodiester bond 3´ & 5´ to AP site
1 nt gap
dR5P OH
Glycosylase & AP lyase YES
Thymine-DNA glycosylase (TDG)  dU or dT opposite dG  N-glycosidic bond  AP site  N/A  N/A  Glycosylase  No 

 
Table Legend:
3mA 3-methyladenine
5-fU 5-formyluridine
5-hmU 5-hydroxymethyluridine
5-hoU 5-hydroxyuridine
7mG 7-methylguanine
8-oxoG 8-oxo-7,8-dihydroguanine
AP apurinic/apyrimidinic sites
CPDs Cyclobutane pyrimidine dimers
dG  deoxyguanine 
dI deoxyisine
dR5P deoxyribose 5´-phosphate
dT  deoxythymine 
dU deoxyuridine
dX deoxyxanthosine
N/A Not applicable
OH Hydroxyl
P Phosphate
PA 3´-phospho-α, β-unsaturated aldehyde
rN ribonucleotides
Tg Thymine Glycol

Foottes

  1. activity is on dsDNA unless ted otherwise
  2. Mir activities, substrates, and references can be found at www.neb.com
  3. 5´ flaps of 1 - 40 nt in length have been confirmed substrates
  4. enzyme has robust activity on ssDNA in addition to dsDNA
  5. Antarctic Thermolabile UDG can be heat inactivated
  6. CPD still covalently attached