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

Wouter L. De Laat, Esther Appeldoorn, Kaoru Sugasawa, Eric Weterings, Nicolaas G.J. Jaspers, Jan H.J. Hoeijmakers

Research output: Contribution to journalArticlepeer-review

274 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)2598-2609
Number of pages12
JournalGenes and Development
Issue number16
StatePublished - Aug 15 1998


  • DNA- binding
  • Nucleotide excision repair
  • Polarity
  • Replication protein A
  • XPG

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology


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