Journal article
Interactions of human replication protein A with oligonucleotides
Biochemistry (Easton), Vol.33(47), pp.14197-14206
11/01/1994
DOI: 10.1021/bi00251a031
PMID: 7947831
Abstract
Replication protein A (RPA) is a heterotrimeric, single-stranded DNA binding protein that is essential for eukaryotic DNA replication. In order to gain a better understanding of the interactions between RPA and DNA, we have examined the interactions of human RPA with single-stranded oligonucleotides. Our analysis of RPA.DNA complexes demonstrated that RPA binds as a heterotrimer. Stoichiometric binding reactions monitored by fluorescence quenching indicated that the binding site size of human RPA is 30 nucleotides and that between 20-30 nucleotides of DNA directly interact with RPA. The binding of RPA to DNA of different lengths was systematically examined using deoxythymidine-containing oligonucleotides. We found that the binding affinity of RPA for short oligonucleotides was length dependent. The apparent association constant of RPA varied over 200-fold from approximately 7 x 10(7) M-1 for oligo(dT)10 to approximately 1.5 x 10(10) M-1 for oligo(dT)50. Human RPA binds to oligonucleotides with low cooperativity; the cooperativity parameter (omega) for RPA binding was estimated to be approximately 15.
Details
- Title: Subtitle
- Interactions of human replication protein A with oligonucleotides
- Creators
- Changsoo KimBrian F PaulusMarc S Wold
- Resource Type
- Journal article
- Publication Details
- Biochemistry (Easton), Vol.33(47), pp.14197-14206
- Publisher
- American Chemical Society
- DOI
- 10.1021/bi00251a031
- PMID
- 7947831
- ISSN
- 0006-2960
- eISSN
- 1520-4995
- Language
- English
- Date published
- 11/01/1994
- Academic Unit
- Radiation Oncology; Biochemistry and Molecular Biology
- Record Identifier
- 9984024556502771
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