Journal article
Replication Protein A Interactions with DNA. III. Molecular Basis of Recognition of Damaged DNA
Biochemistry (Easton), Vol.39(5), pp.850-859
02/08/2000
DOI: 10.1021/bi991704s
PMID: 10653628
Abstract
Human replication protein A (RPA) is a heterotrimeric single-stranded DNA-binding protein (subunits of 70, 32, and 14 kDa) that is required for cellular DNA metabolism. RPA has been reported to interact specifically with damaged double-stranded DNA and to participate in multiple steps of nucleotide excision repair (NER) including the damage recognition step. We have examined the mechanism of RPA binding to both single-stranded and double-stranded DNA (ssDNA and dsDNA, respectively) containing damage. We show that the affinity of RPA for damaged dsDNA correlated with disruption of the double helix by the damaged bases and required RPAs ssDNA-binding activity. We conclude that RPA is recognizing single-stranded character caused by the damaged nucleotides. We also show that RPA binds specifically to damaged ssDNA. The specificity of binding varies with the type of damage with RPA having up to a 60-fold preference for a pyrimidine(6−4)pyrimidone photoproduct. We show that this specific binding was absolutely dependent on the zinc-finger domain in the C-terminus of the 70-kDa subunit. The affinity of RPA for damaged ssDNA was 5 orders of magnitude higher than that of the damage recognition protein XPA (xeroderma pigmentosum group A protein). These findings suggest that RPA probably binds to both damaged and undamaged strands in the NER excision complex. RPA binding may be important for efficient excision of damaged DNA in NER.
Details
- Title: Subtitle
- Replication Protein A Interactions with DNA. III. Molecular Basis of Recognition of Damaged DNA
- Creators
- Ye LaoXavier V GomesYingjie RenJohn-Stephen TaylorMarc S Wold
- Resource Type
- Journal article
- Publication Details
- Biochemistry (Easton), Vol.39(5), pp.850-859
- Publisher
- American Chemical Society
- DOI
- 10.1021/bi991704s
- PMID
- 10653628
- ISSN
- 0006-2960
- eISSN
- 1520-4995
- Language
- English
- Date published
- 02/08/2000
- Academic Unit
- Radiation Oncology; Biochemistry and Molecular Biology
- Record Identifier
- 9984025253502771
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