title: 2-Hydrazinobenzothiazole-based etheno-adduct repair protocol (HERP): a method for quantitative determination of direct repair of etheno-bases.
abstract: Etheno-DNA adducts are mutagenic and lead to genomic instability. Enzymes belonging to Fe(II)/2-oxoglutarate-dependent dioxygenase family repair etheno-DNA adducts by directly removing alkyl chain as glyoxal. Presently there is no simple method to assess repair reaction of etheno-adducts. We have developed a rapid and sensitive assay for studying etheno-DNA adduct repair by Fe(II)/2-oxoglutarate-dependent dioxygenases. Using AlkB as model Fe(II)/2-oxoglutarate-dependent dioxygenases, we performed in vitro repair of etheno-adducts containing DNA and detected glyoxal by reacting with 2-hydrazinobenzothiazole which forms complex yellow color compound with distinct absorption spectrum with a peak absorption at 365 nm. We refer this method as 2-hydrazinobenzothiazole-based etheno-adduct repair protocol or HERP. Our novel approach for determining repair of etheno-adducts containing DNA overcomes several drawbacks of currently available radioisotope-based assay.
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Hyderabad, Ordnance Factory Estate, Yeddumailaram, 502205 Hyderabad, India.
authors.forename: Gururaj
authors.initials: G
authors.lastname: Shivange
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Hyderabad, Ordnance Factory Estate, Yeddumailaram, 502205 Hyderabad, India.
authors.forename: Naveena
authors.initials: N
authors.lastname: Kodipelli
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Hyderabad, Ordnance Factory Estate, Yeddumailaram, 502205 Hyderabad, India. Electronic address: anindya@iith.ac.in.
authors.forename: Roy
authors.initials: R
authors.lastname: Anindya
mesh_terms: D004247:DNA / Q000378:metabolism; D018736:DNA Adducts / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D004260:DNA Repair; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000378:metabolism*; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D013053:Spectrophotometry / Q000379:methods*; D013844:Thiazoles / Q000737:chemistry*
chemical_list: D018736:DNA Adducts; D029968:Escherichia coli Proteins; D013844:Thiazoles; C057550:2-hydrazinobenzothiazole; D004247:DNA; D006899:Mixed Function Oxygenases; C466427:AlkB protein, E coli
keywords: 1,N(2)-ethenoguanine; 1,N(6)-ethenoadenine; 3,N(4)-ethenocytosine; Chloroacetaldehyde; DNA repair; Etheno adduct; Fe(II)/2-oxoglutarate-dependent dioxygenase; N(2),3-ethenoguanine