Journal article
Contributions of the RAD51 N-terminal domain to BRCA2-RAD51 interaction
Nucleic acids research, Vol.41(19), pp.9020-9032
10/2013
DOI: 10.1093/nar/gkt691
PMCID: PMC3799448
PMID: 23935068
Abstract
RAD51 DNA strand exchange protein catalyzes the central step in homologous recombination, a cellular process fundamentally important for accurate repair of damaged chromosomes, preservation of the genetic integrity, restart of collapsed replication forks and telomere maintenance. BRCA2 protein, a product of the breast cancer susceptibility gene, is a key recombination mediator that interacts with RAD51 and facilitates RAD51 nucleoprotein filament formation on single-stranded DNA generated at the sites of DNA damage. An accurate atomistic level description of this interaction, however, is limited to a partial crystal structure of the RAD51 core fused to BRC4 peptide. Here, by integrating homology modeling and molecular dynamics, we generated a structure of the full-length RAD51 in complex with BRC4 peptide. Our model predicted previously unknown hydrogen bonding patterns involving the N-terminal domain (NTD) of RAD51. These interactions guide positioning of the BRC4 peptide within a cavity between the core and the NTDs; the peptide binding separates the two domains and restricts internal dynamics of RAD51 protomers. The model’s depiction of the RAD51-BRC4 complex was validated by free energy calculations and
in vitro
functional analysis of rationally designed mutants. All generated mutants, RAD51
E42A
, RAD51
E59A
, RAD51
E237A
, RAD51
E59A/E237A
and RAD51
E42A/E59A/E237A
maintained basic biochemical activities of the wild-type RAD51, but displayed reduced affinities for the BRC4 peptide. Strong correlation between the calculated and experimental binding energies confirmed the predicted structure of the RAD51-BRC4 complex and highlighted the importance of RAD51 NTD in RAD51-BRCA2 interaction.
Details
- Title: Subtitle
- Contributions of the RAD51 N-terminal domain to BRCA2-RAD51 interaction
- Creators
- Shyamal Subramanyam - Department of Biochemistry, University of Illinois Urbana-Champaign, Urbana, IL 61801, USAWilliam T Jones - Department of Biochemistry, University of Illinois Urbana-Champaign, Urbana, IL 61801, USAMaria Spies - Department of Biochemistry, University of Illinois Urbana-Champaign, Urbana, IL 61801, USAM Ashley Spies - Department of Biochemistry, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
- Resource Type
- Journal article
- Publication Details
- Nucleic acids research, Vol.41(19), pp.9020-9032
- Publisher
- Oxford University Press
- DOI
- 10.1093/nar/gkt691
- PMID
- 23935068
- PMCID
- PMC3799448
- ISSN
- 0305-1048
- eISSN
- 1362-4962
- Language
- English
- Date published
- 10/2013
- Academic Unit
- Pharmaceutical Sciences and Experimental Therapeutics; Radiation Oncology; Biochemistry and Molecular Biology; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984024518202771
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