4OU0

Crystal Structure of RPA32C

Summary for 4OU0

• Entry DOI: 10.2210/pdb4ou0/pdb
• Related: 1DPU
• Descriptor: Replication protein A 32 kDa subunit (2 entities in total)
• Functional Keywords: winged-helix turn helix, protein-protein interaction, s-methyl-thio-cysteine, dna binding protein
• Biological source: Homo sapiens (human)
• Cellular location: Nucleus : P15927
• Total number of polymer chains: 1
• Total formula weight: 7995.88

Authors

Feldkamp, M.D.,Mason, A.C.,Eichman, B.F.,Chazin, W.J. (deposition date: 2014-02-14, release date: 2014-04-30, Last modification date: 2023-09-20)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.4 Å)