4OU0

Crystal Structure of RPA32C

4OU0 の概要

• エントリーDOI: 10.2210/pdb4ou0/pdb
• 関連するPDBエントリー: 1DPU
• 分子名称: Replication protein A 32 kDa subunit (2 entities in total)
• 機能のキーワード: winged-helix turn helix, protein-protein interaction, s-methyl-thio-cysteine, dna binding protein
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Nucleus : P15927
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 7995.88

構造登録者

Feldkamp, M.D.,Mason, A.C.,Eichman, B.F.,Chazin, W.J. (登録日: 2014-02-14, 公開日: 2014-04-30, 最終更新日: 2023-09-20)

主引用文献

Feldkamp, M.D.,Mason, A.C.,Eichman, B.F.,Chazin, W.J.
Structural Analysis of Replication Protein A Recruitment of the DNA Damage Response Protein SMARCAL1.
Biochemistry, 53:3052-3061, 2014

PubMed Abstract: SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily A-like1 (SMARCAL1) is a recently identified DNA damage response protein involved in remodeling stalled replication forks. The eukaryotic single-strand DNA binding protein replication protein A (RPA) recruits SMARCAL1 to stalled forks in vivo and facilitates regression of forks containing leading strand gaps. Both activities are mediated by a direct interaction between an RPA binding motif (RBM) at the N-terminus of SMARCAL1 and the C-terminal winged-helix domain of the RPA 32 kDa subunit (RPA32C). Here we report a biophysical and structural characterization of the SMARCAL1-RPA interaction. Isothermal titration calorimetry and circular dichroism spectroscopy revealed that RPA32C binds SMARCAL1-RBM with a Kd of 2.5 μM and induces a disorder-to-helix transition. The crystal structure of RPA32C was refined to 1.4 Å resolution, and the SMARCAL1-RBM binding site was mapped on the structure on the basis of nuclear magnetic resonance chemical shift perturbations. Conservation of the interaction surface to other RBM-containing proteins allowed construction of a model for the RPA32C/SMARCAL1-RBM complex. The implications of our results are discussed with respect to the recruitment of SMARCAL1 and other DNA damage response and repair proteins to stalled replication forks.

PubMed: 24730652
DOI: 10.1021/bi500252w

実験手法

X-RAY DIFFRACTION (1.4 Å)