RPA stabilizes the XPA-damaged DNA complex through protein-protein interaction

Mu Wang, Alan Mahrenholz, Suk Hee Lee

Research output: Contribution to journalArticle

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Abstract

The xeroderma pigmentosum group A complementing protein (XPA) and eukaryotic replication protein A (RPA) are among the major damage-recognition proteins involved in the early stage of nucleotide excision repair (NER). XPA and RPA are able to bind damaged DNA independently, although RPA interaction stimulates XPA binding to damaged DNA [Li, L., Lu, X., Peterson, C. A., and Legerski, R. J. (1995) Mol. Cell. Biol. 15, 5396-5402 (I); Stigger, E., Drissi, R., and Lee, S.-H. (1998) J. Biol. Chem. 273, 9337-9343 (2)]. In this study, we used surface plasmon resonance (SPR) analysis to investigate the interaction of XPA and RPA with two major types of UV-damaged DNA: the (6-4) photoproduct and the cis-syn cyclobutane dimer of thymidine. Both XPA and RPA preferentially bind to (6-4) photoproduct-containing duplex DNA over cis-syn cyclobutane dimer-containing DNA. The binding of XPA to (6-4) photoproduct was weak (K(D) = 2.13 x 10-8 M), whereas RPA showed a very stable interaction with (6-4) photoproduct (K(D) = 2.02 x 10-10 M). When XPA and RPA were incubated together, the stability of the XPA-damaged DNA interaction was significantly enhanced by wild-type RPA. On the other hand, mutant RPA (RPA:p34Δ33C) defective in its interaction with XPA failed to stabilize XPA- damaged DNA complex. Taken together, our results suggest that a role for RPA in UV-damage recognition is to stabilize XPA-damaged DNA complex through protein-protein interaction.

Original languageEnglish (US)
Pages (from-to)6433-6439
Number of pages7
JournalBiochemistry
Volume39
Issue number21
DOIs
StatePublished - May 30 2000

Fingerprint

Replication Protein A
DNA
Proteins
Cyclobutanes
Dimers
Xeroderma Pigmentosum Group A Protein
Surface Plasmon Resonance
Surface plasmon resonance
Mutant Proteins
DNA Repair
Thymidine

ASJC Scopus subject areas

  • Biochemistry

Cite this

RPA stabilizes the XPA-damaged DNA complex through protein-protein interaction. / Wang, Mu; Mahrenholz, Alan; Lee, Suk Hee.

In: Biochemistry, Vol. 39, No. 21, 30.05.2000, p. 6433-6439.

Research output: Contribution to journalArticle

Wang, Mu ; Mahrenholz, Alan ; Lee, Suk Hee. / RPA stabilizes the XPA-damaged DNA complex through protein-protein interaction. In: Biochemistry. 2000 ; Vol. 39, No. 21. pp. 6433-6439.
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AB - The xeroderma pigmentosum group A complementing protein (XPA) and eukaryotic replication protein A (RPA) are among the major damage-recognition proteins involved in the early stage of nucleotide excision repair (NER). XPA and RPA are able to bind damaged DNA independently, although RPA interaction stimulates XPA binding to damaged DNA [Li, L., Lu, X., Peterson, C. A., and Legerski, R. J. (1995) Mol. Cell. Biol. 15, 5396-5402 (I); Stigger, E., Drissi, R., and Lee, S.-H. (1998) J. Biol. Chem. 273, 9337-9343 (2)]. In this study, we used surface plasmon resonance (SPR) analysis to investigate the interaction of XPA and RPA with two major types of UV-damaged DNA: the (6-4) photoproduct and the cis-syn cyclobutane dimer of thymidine. Both XPA and RPA preferentially bind to (6-4) photoproduct-containing duplex DNA over cis-syn cyclobutane dimer-containing DNA. The binding of XPA to (6-4) photoproduct was weak (K(D) = 2.13 x 10-8 M), whereas RPA showed a very stable interaction with (6-4) photoproduct (K(D) = 2.02 x 10-10 M). When XPA and RPA were incubated together, the stability of the XPA-damaged DNA interaction was significantly enhanced by wild-type RPA. On the other hand, mutant RPA (RPA:p34Δ33C) defective in its interaction with XPA failed to stabilize XPA- damaged DNA complex. Taken together, our results suggest that a role for RPA in UV-damage recognition is to stabilize XPA-damaged DNA complex through protein-protein interaction.

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