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Updated: Mar 18, 2026

Visualization of DNA Repair Proteins Interaction by Immunofluorescence
Published on: June 26, 2020
RecA stimulates AlkB-mediated direct repair of DNA adducts
Gururaj Shivange1, Mohan Monisha1, Richa Nigam1
1Department of Biotechnology, Indian Institute of Technology Hyderabad, Kandi, 502285 Hyderabad, Telangana, India.
Abstract:
The Escherichia coli AlkB protein is a 2-oxoglutarate/Fe(II)-dependent demethylase that repairs alkylated single stranded and double stranded DNA. Immunoaffinity chromatography coupled with mass spectrometry identified RecA, a key factor in homologous recombination, as an AlkB-associated protein. The interaction between AlkB and RecA was validated by yeast two-hybrid assay; size-exclusion chromatography and standard pull down experiment and was shown to be direct and mediated by the N-terminal domain of RecA. RecA binding results AlkB-RecA heterodimer formation and RecA-AlkB repairs alkylated DNA with higher efficiency than AlkB alone.
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