Identification and characterization of a human DNA glycosylase for repair of modified bases in oxidatively damaged DNA

Tapas K. Hazra, Tadahide Izumi, Istvan Boldogh, Barry Imhoff, Yoke W. Kow, Pawel Jaruga, Miral Dizdaroglu, Sankar Mitra

Research output: Contribution to journalArticlepeer-review

459 Scopus citations

Abstract

8-Oxoguanine (8-oxoG), ring-opened purines (formamidopyrimidines or Fapys), and other oxidized DNA base lesions generated by reactive oxygen species are often mutagenic and toxic, and have been implicated in the etiology of many diseases, including cancer, and in aging. Repair of these lesions in all organisms occurs primarily via the DNA base excision repair pathway, initiated with their excision by DNA glycosylase/AP lyases, which are of two classes. One class utilizes an internal Lys residue as the active site nucleophile, and includes Escherichia coli Nth and both known mammalian DNA glycosylase/AP lyases, namely, OGG1 and NTH1. E. coli MutM and its paralog Nei, which comprise the second class, use N-terminal Pro as the active site. Here, we report the presence of two human orthologs of E. coli mutM nei genes in the human genome database, and characterize one of their products. Based on the substrate preference, we have named it NEH1 (Nei homolog). The 44-kDa, wild-type recombinant NEH1, purified to homogeneity from E. coli, excises Fapys from damaged DNA, and oxidized pyrimidines and 8-oxoG from oligodeoxynucleotides. Inactivation of the enzyme because of either deletion of N-terminal Pro or Histag fusion at the N terminus supports the role of N-terminal Pro as its active site. The tissue-specific levels of NEH1 and OGG1 mRNAs are distinct, and S phase-specific increase in NEH1 at both RNA and protein levels suggests that NEH1 is involved in replication-associated repair of oxidized bases.

Original languageEnglish (US)
Pages (from-to)3523-3528
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume99
Issue number6
DOIs
StatePublished - Mar 19 2002

ASJC Scopus subject areas

  • General

Fingerprint

Dive into the research topics of 'Identification and characterization of a human DNA glycosylase for repair of modified bases in oxidatively damaged DNA'. Together they form a unique fingerprint.

Cite this