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Post‐replicative base excision repair in replication foci
Author(s) -
Otterlei Marit,
Warbrick Emma,
Nagelhus Toril A.,
Haug Terje,
Slupphaug Geir,
Akbari Mansour,
Aas Per Arne,
Steinsbekk Kristin,
Bakke Oddmund,
Krokan Hans E.
Publication year - 1999
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/18.13.3834
Subject(s) - biology , replication (statistics) , base excision repair , dna repair , nucleotide excision repair , genetics , dna , virology
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.