Document Type

Article

Peer Reviewed

1

Publication Date

7-8-2016

NLM Title Abbreviation

Nucl. Acids Res.

Journal/Book/Conference Title

Nucleic acids research

PubMed ID

27131385

DOI of Published Version

10.1093/nar/gkw339

Abstract

Replication protein A (RPA), the major eukaryotic single-stranded DNA (ssDNA) binding protein, is essential for replication, repair and recombination. High-affinity ssDNA-binding by RPA depends on two DNA binding domains in the large subunit of RPA. Mutation of the evolutionarily conserved aromatic residues in these two domains results in a separation-of-function phenotype: aromatic residue mutants support DNA replication but are defective in DNA repair. We used biochemical and single-molecule analyses, and Brownian Dynamics simulations to determine the molecular basis of this phenotype. Our studies demonstrated that RPA binds to ssDNA in at least two modes characterized by different dissociation kinetics. We also showed that the aromatic residues contribute to the formation of the longer-lived state, are required for stable binding to short ssDNA regions and are needed for RPA melting of partially duplex DNA structures. We conclude that stable binding and/or the melting of secondary DNA structures by RPA is required for DNA repair, including RAD51 mediated DNA strand exchange, but is dispensable for DNA replication. It is likely that the binding modes are in equilibrium and reflect dynamics in the RPA-DNA complex. This suggests that dynamic binding of RPA to DNA is necessary for different cellular functions.

Keywords

OAfund

Journal Article Version

Version of Record

Published Article/Book Citation

Nucl. Acids Res. (08 July 2016) 44 (12): 5758-5772. doi: 10.1093/nar/gkw339

Rights

© The Author(s) 2016

Creative Commons License

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.

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URL

https://ir.uiowa.edu/biochem_pubs/1