Formamidopyrimidine-DNA glycosylase of Escherichia coli: cloning and sequencing of the fpg structural gene and overproduction of the protein.

Boiteux, Serge; O’Connor, Timothy; Laval, Jacques

doi:10.1002/j.1460-2075.1987.tb02629.x

Cited by 287 publications

(166 citation statements)

References 37 publications

Supporting

Mentioning

165

Contrasting

Order By: Relevance

“…The fpg gene encodes a protein of 30.2 kDa [210,211]. In addition to its N-glycosylase activity, this protein also has a nicking activity that cleaves both the 5h-and 3h-phosphodiester bonds at an AP site by successive β-and δ-elimination reactions, leaving both the 3h and 5h DNA ends phosphorylated [212].…”

Section: Fpg Family and The Oxog Systemmentioning

confidence: 99%

DNA glycosylases in the base excision repair of DNA

Krokan

Standal

Slupphaug

1997

Biochemical Journal

771

566

View full text Add to dashboard Cite

A wide range of cytotoxic and mutagenic DNA bases are removed by different DNA glycosylases, which initiate the base excision repair pathway. DNA glycosylases cleave the Nglycosylic bond between the target base and deoxyribose, thus releasing a free base and leaving an apurinic\apyrimidinic (AP) site. In addition, several DNA glycosylases are bifunctional, since they also display a lyase activity that cleaves the phosphodiester backbone 3h to the AP site generated by the glycosylase activity. Structural data and sequence comparisons have identified common features among many of the DNA glycosylases. Their active sites have a structure that can only bind extrahelical target bases, as observed in the crystal structure of human uracil-DNA glycosylase in a complex with double-stranded DNA. Nucleotide flipping is apparently actively facilitated by the enzyme. With bacteriophage T4 endonuclease V, a pyrimidine-

show abstract

Section: Fpg Family and The Oxog Systemmentioning

confidence: 99%

DNA glycosylases in the base excision repair of DNA

Krokan

Standal

Slupphaug

1997

Biochemical Journal

771

566

View full text Add to dashboard Cite

show abstract

“…The cells were harvested by centrifugation, washed, and stored at Ϫ80°C. Cell-free extracts were prepared as previously described (32), and protein concentration was measured according to Bradford (33).…”

Section: Strainsmentioning

confidence: 99%

Cloning and characterization of hOGG1 , a human homolog of the OGG1 gene of Saccharomyces cerevisiae

Radicella

Dhérin

Desmaze

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

565

305

View full text Add to dashboard Cite

The OGG1 gene of Saccharomyces cerevisiae encodes a DNA glycosylase activity that is a functional analog of the Fpg protein from Escherichia coli and excises 7,8-dihydro-8-oxoguanine (8-oxoG) from damaged DNA. The repair of this ubiquitous kind of oxidative damage is essential to prevent mutations both in bacteria and in yeast. A human cDNA clone carrying an ORF displaying homology to the yeast protein was identified. The predicted protein has 345 amino acids and a molecular mass of 39 kDa. This protein shares a 38% sequence identity with the yeast Ogg1 protein, adding this novel human gene product to the growing family of enzymes that the repair of oxidatively damaged bases and are related to the E. coli endonuclease III. Northern blot analysis indicates that this gene, localized to chromosome 3p25, is ubiquitously expressed in human tissues. The cloned coding sequence was expressed in an E. coli strain that carried a disrupted fpg gene, the bacterial functional analog of OGG1. Cell-free extracts from these cultures displayed a specific lyase activity on duplex DNA that carried an 8-oxoG͞C base pair. The products of the reaction are consistent with an enzymatic activity like the one displayed by the yeast Ogg1. Analysis of the substrate specificity reveals a very strong preference for DNA fragments harboring 8-oxoG͞C base pairs. The pattern of specificity correlates well with the one found for the yeast enzyme. Moreover, when the human coding sequence was expressed in a yeast strain mutant in OGG1 it was able to complement the spontaneous mutator phenotype. These results make this novel gene (hOGG1) a strong candidate for the human homolog of the yeast OGG1 and suggest an important role of its product in the protection of the genome from the mutagenic effects of the oxidatively damaged purines.Reactive oxygen species (ROS) formed in cells either as by-products of aerobic metabolism or as a consequence of exposure to environmental mutagens can attack DNA or its precursors, yielding oxidatively damaged bases and strand breakage (1, 2). Unrepaired oxidative damage to DNA has been suggested to play a role in carcinogenesis and aging through mutations in genes controlling these biological processes (3-5). Several lines of evidence suggest that an oxidatively damaged form of guanine, 7,8-dihydro-8-oxoguanine (8-oxoG), is critical in terms of mutagenesis (6, 7). In Escherichia coli, two DNA glycosylases cooperate to prevent mutagenesis by 8-oxoG: the Fpg protein, which excises 8-oxoG in damaged DNA (8-10) and the MutY protein, which excises the adenine residues incorporated by DNA polymerases opposite 8-oxoG (11-13). Inactivation of both the fpg (mutM) and mutY (micA) genes of E. coli results in a strong G⅐C 3 T⅐A mutator phenotype (14-17).In Saccharomyces cerevisiae, the OGG1 gene encodes an 8-oxoG DNA glycosylase activity that reduces the mutator phenotype of the fpg mutY mutant of E. coli (18). The Ogg1 protein contains 376 amino acids, and, although it was cloned by functional complementation of the E. co...

show abstract

“…These ring-opened purines are excised from DNA by a DNA glycosylase, formamido-pyrimidine-DNA glycosylase (Chetsanga & Lindahl, 1979;Breimer, 1984) which is also present in mammalian cells (Breimer, 1983). The gene of the E. coli enzyme has now been cloned and sequenced (Boiteux et al, 1987). Thus its physiological role should soon be known.…”

Section: Dna Lesions and Their Repairmentioning

confidence: 99%