DNA-binding polarity of human replication protein A positions nucleases in nucleotide excision repair

The human single-stranded DNA-binding replication A protein (RPA) is involved in various DNA-processing events. By comparing the affinity of hRPA for artificial DNA hairpin structures with 3'- or 5'-protruding single-stranded arms, we found that hRPA binds ssDNA with a defined polarity; a...

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Bibliographic Details
Published in:Genes & development 1998-08, Vol.12 (16), p.2598-2609
Main Authors: de Laat, W L, Appeldoorn, E, Sugasawa, K, Weterings, E, Jaspers, N G, Hoeijmakers, J H
Format: Article
Language:English
Subjects:
Animals
Cells, Cultured
DNA - metabolism
DNA Repair
DNA, Single-Stranded - metabolism
DNA-Binding Proteins - metabolism
Endonucleases - metabolism
Humans
Insecta
Nuclear Proteins
Protein Binding
Proteins - metabolism
Replication Protein A
Research Paper
Substrate Specificity
Transcription Factors
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Summary:The human single-stranded DNA-binding replication A protein (RPA) is involved in various DNA-processing events. By comparing the affinity of hRPA for artificial DNA hairpin structures with 3'- or 5'-protruding single-stranded arms, we found that hRPA binds ssDNA with a defined polarity; a strong ssDNA interaction domain of hRPA is positioned at the 5' side of its binding region, a weak ssDNA-binding domain resides at the 3' side. Polarity appears crucial for positioning of the excision repair nucleases XPG and ERCC1-XPF on the DNA. With the 3'-oriented side of hRPA facing a duplex ssDNA junction, hRPA interacts with and stimulates ERCC1-XPF, whereas the 5'-oriented side of hRPA at a DNA junction allows stable binding of XPG to hRPA. Our data pinpoint hRPA to the undamaged strand during nucleotide excision repair. Polarity of hRPA on ssDNA is likely to contribute to the directionality of other hRPA-dependent processes as well.
ISSN:0890-9369
1549-5477
DOI:10.1101/gad.12.16.2598