1AA3

C-TERMINAL DOMAIN OF THE E. COLI RECA, NMR, MINIMIZED AVERAGE STRUCTURE

Summary for 1AA3

• Entry DOI: 10.2210/pdb1aa3/pdb
• Descriptor: RECA (1 entity in total)
• Functional Keywords: double-stranded dna binding domain, riken structural genomics/proteomics initiative, rsgi, structural genomics
• Biological source: Escherichia coli
• Cellular location: Cytoplasm: P0A7G6
• Total number of polymer chains: 1
• Total formula weight: 7108.13

Authors

Aihara, H.,Ito, Y.,Kurumizaka, H.,Terada, T.,Yokoyama, S.,Shibata, T.,RIKEN Structural Genomics/Proteomics Initiative (RSGI) (deposition date: 1997-01-22, release date: 1997-07-23, Last modification date: 2024-04-10)

Primary citation

Aihara, H.,Ito, Y.,Kurumizaka, H.,Terada, T.,Yokoyama, S.,Shibata, T.
An interaction between a specified surface of the C-terminal domain of RecA protein and double-stranded DNA for homologous pairing.
J.Mol.Biol., 274:213-221, 1997

PubMed Abstract: RecA protein and its homologs catalyze homologous pairing of dsDNA and ssDNA, a critical reaction in homologous genetic recombination in various organisms from a virus, microbes to higher eukaryotes. In this reaction, RecA protein forms a nucleoprotein filament on ssDNA, which in turn binds to naked dsDNA for homology search. We suggested that the C-terminal domain of RecA protein plays a role in capturing the dsDNA. Here, we isolated the C-terminal domain as a soluble form and determined the solution structure by NMR spectroscopy. The overall folding of the NMR structure agrees with that of the corresponding part of the reported crystal structure, but a remarkable difference was found in a solvent-exposed region due to intermolecular contacts in the crystal. Then, we studied the interaction between the C-terminal domain and DNA, and found that significant chemical shift changes were induced in a specific region by titration with dsDNA. SsDNA induced a much smaller chemical shift perturbation. The difference of DNA concentrations to give the half-saturation of the chemical shift change showed a higher affinity of the C-terminal region toward dsDNA. Combined with our previous results, these provide direct evidence that the defined region in the C-terminal domain furnishes a binding surface for DNA.

PubMed: 9398528
DOI: 10.1006/jmbi.1997.1403

Experimental method

SOLUTION NMR