Journal article
Role of Protein−Protein Interactions in the Function of Replication Protein A (RPA): RPA Modulates the Activity of DNA Polymerase α by Multiple Mechanisms
Biochemistry (Easton), Vol.36(28), pp.8443-8454
07/15/1997
DOI: 10.1021/bi970473r
PMID: 9214288
Abstract
Replication Protein A (RPA) from human cells is a stable complex of 70-, 32-, and 14-kDa subunits that is required for multiple processes in DNA metabolism. RPA binds with high affinity to single-stranded DNA and interacts with multiple proteins, including proteins required for the initiation of SV40 DNA replication, DNA polymerase α and SV40 large T antigen. We have used a series of mutant derivatives of RPA to map the regions of RPA required for specific protein−protein interactions and have examined the roles of these interactions in DNA replication. T antigen, DNA polymerase α and the activation domain of VP16 all have overlapping sites of interaction in the N-terminal half (residues 1−327) of the 70-kDa subunit of RPA. In addition, the interaction site for DNA polymerase α is composed of two functionally distinct regions, one (residues 1−∼170) which stimulates polymerase activity and a second (residues ∼170−327) which increases polymerase processivity. In the latter, both the direct protein−protein interaction and ssDNA-binding activities of RPA were needed for RPA to modulate polymerase processivity. We also found that SV40 T antigen inhibited the ability of RPA to increase processivity of DNA polymerase α, suggesting that this activity of RPA may be important for elongation but not during the initiation of DNA replication. DNA polymerase α, but not T antigen also interacted with the 32- and/or 14-kDa subunits of RPA, but these interactions did not seem to effect polymerase activity.
Details
- Title: Subtitle
- Role of Protein−Protein Interactions in the Function of Replication Protein A (RPA): RPA Modulates the Activity of DNA Polymerase α by Multiple Mechanisms
- Creators
- Katherine A BraunYe LaoZhigang HeC. James InglesMarc S Wold
- Resource Type
- Journal article
- Publication Details
- Biochemistry (Easton), Vol.36(28), pp.8443-8454
- Publisher
- American Chemical Society
- DOI
- 10.1021/bi970473r
- PMID
- 9214288
- ISSN
- 0006-2960
- eISSN
- 1520-4995
- Language
- English
- Date published
- 07/15/1997
- Academic Unit
- Radiation Oncology; Biochemistry and Molecular Biology
- Record Identifier
- 9984024526902771
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