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Post-replicative base excision repair in replication foci
- Publication Year :
- 1999
-
Abstract
- 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 uracil-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 uracil-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.
- Subjects :
- DNA Replication
DNA Repair
Base Pair Mismatch
Recombinant Fusion Proteins
Molecular Sequence Data
Gene Expression
Biology
DNA polymerase delta
General Biochemistry, Genetics and Molecular Biology
Cell Line
DNA Glycosylases
Replication factor C
Control of chromosome duplication
Proliferating Cell Nuclear Antigen
Replication Protein A
Yeasts
Humans
Amino Acid Sequence
Uracil
Uracil-DNA Glycosidase
Molecular Biology
Replication protein A
N-Glycosyl Hydrolases
Cell Nucleus
Binding Sites
General Immunology and Microbiology
General Neuroscience
Cell Cycle
DNA replication
Base excision repair
DNA
Molecular biology
Peptide Fragments
DNA-Binding Proteins
Kinetics
DNA glycosylase
Uracil-DNA glycosylase
Deoxyuracil Nucleotides
Research Article
HeLa Cells
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....6fafea0c820444e73d9825ddb522dadd