3LDA

Yeast Rad51 H352Y Filament Interface Mutant

3LDA の概要

• エントリーDOI: 10.2210/pdb3lda/pdb
• 関連するPDBエントリー: 1SZP
• 分子名称: DNA repair protein RAD51, CHLORIDE ION (3 entities in total)
• 機能のキーワード: dna binding protein, atp-binding, dna damage, dna recombination, dna repair, nucleotide-binding, non-prolyl cis peptide, atp-hydrolysis, walker a/b
• 由来する生物種: Saccharomyces cerevisiae (yeast)
• 細胞内の位置: Nucleus: P25454
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 43138.71

構造登録者

Villanueva, N.L.,Chen, J.,Morrical, S.W.,Rould, M.A. (登録日: 2010-01-12, 公開日: 2010-04-21, 最終更新日: 2023-09-06)

主引用文献

Chen, J.,Villanueva, N.,Rould, M.A.,Morrical, S.W.
Insights into the mechanism of Rad51 recombinase from the structure and properties of a filament interface mutant.
Nucleic Acids Res., 38:4889-4906, 2010

PubMed Abstract: Rad51 protein promotes homologous recombination in eukaryotes. Recombination activities are activated by Rad51 filament assembly on ssDNA. Previous studies of yeast Rad51 showed that His352 occupies an important position at the filament interface, where it could relay signals between subunits and active sites. To investigate, we characterized yeast Rad51 H352A and H352Y mutants, and solved the structure of H352Y. H352A forms catalytically competent but salt-labile complexes on ssDNA. In contrast, H352Y forms salt-resistant complexes on ssDNA, but is defective in nucleotide exchange, RPA displacement and strand exchange with full-length DNA substrates. The 2.5 A crystal structure of H352Y reveals a right-handed helical filament in a high-pitch (130 A) conformation with P6(1) symmetry. The catalytic core and dimer interface regions of H352Y closely resemble those of DNA-bound Escherichia coli RecA protein. The H352Y mutation stabilizes Phe187 from the adjacent subunit in a position that interferes with the gamma-phosphate-binding site of the Walker A motif/P-loop, potentially explaining the limited catalysis observed. Comparison of Rad51 H352Y, RecA-DNA and related structures reveals that the presence of bound DNA correlates with the isomerization of a conserved cis peptide near Walker B to the trans configuration, which appears to prime the catalytic glutamate residue for ATP hydrolysis.

PubMed: 20371520
DOI: 10.1093/nar/gkq209

実験手法

X-RAY DIFFRACTION (2.5 Å)