Post-replicative base excision repair in replication foci

Base excision repair (BER) is initiated by a DNA glycosylase and is completed by alternative routes, one of which requires proliferating cell nuclear antigen (PCNA) and other proteins also involved in DNA replication. We report that the major nuclear uraci‐DNA glycosylase (UNG2) increases in S phase...

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Bibliographic Details
Published in:The EMBO journal 1999-07, Vol.18 (13), p.3834-3844
Main Authors: Otterlei, Marit, Warbrick, Emma, Nagelhus, Toril A., Haug, Terje, Slupphaug, Geir, Akbari, Mansour, Aas, Per Arne, Steinsbekk, Kristin, Bakke, Oddmund, Krokan, Hans E.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Base Pair Mismatch - genetics
Binding Sites
Cell Cycle
Cell Line
Cell Nucleus - enzymology
Cell Nucleus - metabolism
Deoxyribonucleic acid
Deoxyuracil Nucleotides - metabolism
DNA
DNA - biosynthesis
DNA Glycosylases
DNA Repair - genetics
DNA Replication - genetics
DNA-Binding Proteins - metabolism
Gene Expression
HeLa Cells
Humans
Immunoassays
Kinetics
Molecular Sequence Data
N-Glycosyl Hydrolases - genetics
N-Glycosyl Hydrolases - metabolism
Peptide Fragments - genetics
Peptide Fragments - metabolism
proliferating cell nuclear antigen
Proliferating Cell Nuclear Antigen - metabolism
Recombinant Fusion Proteins - metabolism
replication foci
Replication Protein A
uraci-DNA glycosylase
Uracil - metabolism
Uracil-DNA Glycosidase
Yeasts - cytology
Yeasts - genetics
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Description
Summary:Base excision repair (BER) is initiated by a DNA glycosylase and is completed by alternative routes, one of which requires proliferating cell nuclear antigen (PCNA) and other proteins also involved in DNA replication. We report that the major nuclear uraci‐DNA glycosylase (UNG2) increases in S phase, during which it co‐localizes with incorporated BrdUrd in replication foci. Uracil is rapidly removed from replicatively incorporated dUMP residues in isolated nuclei. Neutralizing antibodies to UNG2 inhibit this removal, indicating that UNG2 is the major uraci‐DNA glycosylase responsible. PCNA and replication protein A (RPA) co‐localize with UNG2 in replication foci, and a direct molecular interaction of UNG2 with PCNA (one binding site) and RPA (two binding sites) was demonstrated using two‐hybrid assays, a peptide SPOT assay and enzyme‐linked immunosorbent assays. These results demonstrate rapid post‐replicative removal of incorporated uracil by UNG2 and indicate the formation of a BER complex that contains UNG2, RPA and PCNA close to the replication fork.
ISSN:0261-4189
1460-2075
1460-2075
DOI:10.1093/emboj/18.13.3834