TY - JOUR
T1 - DNA-binding polarity of human replication protein A positions nucleases in nucleotide excision repair
AU - De Laat, Wouter L.
AU - Appeldoorn, Esther
AU - Sugasawa, Kaoru
AU - Weterings, Eric
AU - Jaspers, Nicolaas G.J.
AU - Hoeijmakers, Jan H.J.
PY - 1998/8/15
Y1 - 1998/8/15
N2 - 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.
AB - 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.
KW - DNA- binding
KW - ERCC1-XPF
KW - Nucleotide excision repair
KW - Polarity
KW - Replication protein A
KW - XPG
UR - http://www.scopus.com/inward/record.url?scp=0032529167&partnerID=8YFLogxK
U2 - 10.1101/gad.12.16.2598
DO - 10.1101/gad.12.16.2598
M3 - Article
C2 - 9716411
AN - SCOPUS:0032529167
SN - 0890-9369
VL - 12
SP - 2598
EP - 2609
JO - Genes and Development
JF - Genes and Development
IS - 16
ER -