Role of zinc-finger motif in redox regulation of human replication protein A

M. Wang, J. S. You, S. H. Lee

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Replication protein A (RPA) is a heterotrimeric zinc-finger protein complex involved in DNA replication, repair, and genetic recombination. Unlike other zinc-finger proteins, RPA's zinc-finger motif is not essential for its single-stranded DNA (ssDNA) binding activity, but is involved in redox regulation of its single-stranded DNA (ss-DNA) binding activity. To get an insight into the regulation of RPA-ssDNA interaction, wild-type RPA (wt-RPA) and zinc-finger mutant were examined for ssDNA binding activity using surface plasmon resonance technique. Interaction of wt-RPA with ssDNA under nonreducing conditions was very weak (KD × 2.33 × 10-8 M) compared with that under reducing conditions (KD = 7.35 × 10-11 M), whereas ssDNA binding affinity of the zinc-finger mutant was not affected by redox. The divalent ion chelator, o-phenanthroline, significantly reduced wt-RPA-ss-DNA interaction, but had no effect on the zinc-finger mutant. The inhibitory effect of o-phenanthroline on RPA-ss-DNA interaction was reversed by Zn(II), but not by other divalent cations, suggesting that Zn(II) is the unique metal coordinating the zinc-finger cysteines in redox regulation of RPA-ssDNA interaction. In DNA repair, redox affected RPA's interaction with damaged DNA, but not its role in stabilizing the xeroderma pigmentosum group A (XPA)-damaged DNA complex, suggesting that the zinc-finger motif may mediate the transition of RPA-XPA interaction to a stable RPA-XPA-damaged DNA complex in a redox-dependent manner.

Original languageEnglish (US)
Pages (from-to)657-669
Number of pages13
JournalAntioxidants and Redox Signaling
Volume3
Issue number4
DOIs
StatePublished - Jan 1 2001

Fingerprint

Replication Protein A
Single-Stranded DNA
Zinc Fingers
Oxidation-Reduction
Zinc
Xeroderma Pigmentosum
DNA
DNA Repair
Repair
human RPA1 protein
Surface Plasmon Resonance
Divalent Cations
Surface plasmon resonance
Chelating Agents
DNA Replication
Genetic Recombination
Cysteine
Proteins
Metals
Ions

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Role of zinc-finger motif in redox regulation of human replication protein A. / Wang, M.; You, J. S.; Lee, S. H.

In: Antioxidants and Redox Signaling, Vol. 3, No. 4, 01.01.2001, p. 657-669.

Research output: Contribution to journalArticle

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