Reference: Rieger RA, et al. (2006) Proteomic approach to identification of proteins reactive for abasic sites in DNA. Mol Cell Proteomics 5(5):858-67

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Abstract


Abasic (AP) sites, a prominent type of DNA damage, are repaired through base excision repair (BER) mechanism in both prokaryotes and eukaryotes and may interfere with many other cellular processes. A full repertoire of AP site-binding proteins in cells is presently unknown, preventing reliable assessment of harm inflicted by these ubiquitous lesions and of their involvement in the flux of DNA metabolism. We present a proteomic-based strategy for assembling at least a partial catalogue of proteins capable of binding AP sites in DNA. The general scheme relies on the sensitivity of many AP site-bound protein species to NaBH4 cross-linking. An affinity-tagged substrate is used to facilitate isolation of the cross-linked species, which are then separated and analyzed by mass spectrometry methods. We report identification of seven proteins from E. coli (AroF, DnaK, MutM, PolA, TnaA, TufA, UvrA) and two proteins from baker's yeast (ARC1, Ygl245wp) reactive for AP sites in this system.

Reference Type
Journal Article
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Rieger RA, Zaika EI, Xie W, Johnson F, Grollman AP, Iden CR, Zharkov DO
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