Full-length archaeal Rad51 structure and mutants: mechanisms for RAD51 assembly and control by BRCA2

Autor: Luca Pellegrini, Chiharu Hitomi, David S. Yu, Mahmud K.K. Shivji, John A. Tainer, Lisa Craig, David S. Shin, Andrew S. Arvai, Tom L. Blundell, Niels Volkmann, Hiro Tsuruta, Douglas S. Daniels, Debbie Bates, Ashok R. Venkitaraman, Biana Yelent
Rok vydání: 2003
Předmět:
Models
Molecular
DNA repair
Archaeal Proteins
Protein subunit
Molecular Sequence Data
genetic processes
RAD52
RAD51
Biology
Crystallography
X-Ray
Models
Biological
General Biochemistry
Genetics and Molecular Biology
Cell Line
Protein–protein interaction
chemistry.chemical_compound
Protein structure
Humans
Amino Acid Sequence
Protein Structure
Quaternary
Molecular Biology
Adenosine Triphosphatases
BRCA2 Protein
Genetics
Binding Sites
Sequence Homology
Amino Acid
General Immunology and Microbiology
General Neuroscience
DNA
Articles
Recombinant Proteins
Protein Structure
Tertiary
DNA-Binding Proteins
Pyrococcus furiosus
Microscopy
Electron
enzymes and coenzymes (carbohydrates)
chemistry
Mutation
health occupations
Biophysics
Rad51 Recombinase
biological phenomena
cell phenomena
and immunity
Homologous recombination
Zdroj: The EMBO Journal. 22:4566-4576
ISSN: 1460-2075
DOI: 10.1093/emboj/cdg429
Popis: To clarify RAD51 interactions controlling homologous recombination, we report here the crystal structure of the full-length RAD51 homolog from Pyrococcus furiosus. The structure reveals how RAD51 proteins assemble into inactive heptameric rings and active DNA-bound filaments matching three-dimensional electron microscopy reconstructions. A polymerization motif (RAD51-PM) tethers individual subunits together to form assemblies. Subunit interactions support an allosteric 'switch' promoting ATPase activity and DNA binding roles for the N-terminal domain helix-hairpin-helix (HhH) motif. Structural and mutational results characterize RAD51 interactions with the breast cancer susceptibility protein BRCA2 in higher eukaryotes. A designed P.furiosus RAD51 mutant binds BRC repeats and forms BRCA2-dependent nuclear foci in human cells in response to gamma-irradiation-induced DNA damage, similar to human RAD51. These results show that BRCA2 repeats mimic the RAD51-PM and imply analogous RAD51 interactions with RAD52 and RAD54. Both BRCA2 and RAD54 may act as antagonists and chaperones for RAD51 filament assembly by coupling RAD51 interface exchanges with DNA binding. Together, these structural and mutational results support an interface exchange hypothesis for coordinated protein interactions in homologous recombination.
Databáze: OpenAIRE
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