2-Hydrazinobenzothiazole-based etheno-adduct repair protocol (HERP): A method for quantitative determination of direct repair of etheno-bases

Shivange, G and Kodipelli, N and Roy, Anindya (2015) 2-Hydrazinobenzothiazole-based etheno-adduct repair protocol (HERP): A method for quantitative determination of direct repair of etheno-bases. DNA Repair, 28. pp. 8-13. ISSN 1568-7864

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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.

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IITH Creators:

IITH Creators	ORCiD
Roy, Anindya	http://orcid.org/0000-0001-8561-907X

Item Type:

Article

Additional Information:

E. coli HK82 (alkB) cells were kindly provided by Dr. Hans Krokan, Norwegian University of Science and Technology.

Uncontrolled Keywords:

Fe(II)/2-oxoglutarate-dependent dioxygenase; Chloroacetaldehyde; Etheno adduct; DNA repair; 1,N6-ethenoadenine; 3,N4-ethenocytosine; N2,3-ethenoguanine; 1,N2-ethenoguanine

Subjects:

Others > Biotechnology

Divisions:

Department of Biotechnology

Depositing User:

Library Staff

Date Deposited:

26 Feb 2015 11:11

Last Modified:

05 Dec 2017 04:51

URI:

http://raiithold.iith.ac.in/id/eprint/1361