The novel DNA glycosylase, NEIL1, protects mammalian cells from radiation-mediated cell death.
DNA damage mediated by reactive oxygen species generates miscoding and blocking lesions that may lead to mutations or cell death. Base excision repair (BER) constitutes a universal mechanism for removing oxidatively damaged bases and restoring the integrity of genomic DNA. In Escherichia coli, the DNA glycosylases Nei, Fpg, and Nth initiate BER of oxidative lesions; OGG1 and NTH1 proteins fulfill a similar function in mammalian cells. Three human genes, designated NEIL1, NEIL2 and NEIL3, encode proteins that contain sequence homologies to Nei and Fpg. We have cloned the corresponding mouse genes and have overexpressed and purified mNeil1, a DNA glycosylase that efficiently removes a wide spectrum of mutagenic and cytotoxic DNA lesions. These lesions include the two cis-thymineglycol(Tg) stereoisomers, guanine- and adenine-derived formamidopyrimidines, and 5,6-dihydrouracil. Two of these lesions, fapyA and 5S,6R thymine glycol, are not excised by mOgg1 or mNth1. We have also used RNA interference technology to establish embryonic stem cell lines deficient in Neil1 protein and showed them to be sensitive to low levels of gamma-irradiation. The results of these studies suggest that Neil1 is an essential component of base excision repair in mammalian cells; its presence may contribute to the redundant repair capacity observed in Ogg1 -/- and Nth1 -/- mice.
Rosenquist
Thomas A
TA
Department of Pharmacological Sciences, SUNY Stony Brook, Stony Brook, NY 11794, USA. rosenquist@pharm.sunysb.edu
Zaika
Elena
E
Fernandes
Andrea S
AS
Zharkov
Dmitry O
DO
Miller
Holly
H
Grollman
Arthur P
AP
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016923:Cell Death; D003001:Cloning, Molecular; D004249:DNA Damage; D045647:DNA Glycosylases / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D004307:Dose-Response Relationship, Radiation; D004591:Electrophoresis, Polyacrylamide Gel; D004622:Embryo, Mammalian / Q000166:cytology; D004926:Escherichia coli / Q000378:metabolism; D005720:Gamma Rays; D006801:Humans; D007700:Kinetics; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009154:Mutation; D009841:Oligonucleotides / Q000737:chemistry; D010100:Oxygen / Q000378:metabolism; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000378:metabolism; D013237:Stereoisomerism; D013997:Time Factors
D009841:Oligonucleotides; D045647:DNA Glycosylases; C473634:NEIL1 protein, human; D010100:Oxygen