3VW4

Crystal structure of the DNA-binding domain of ColE2-P9 Rep in complex with the replication origin

Summary for 3VW4

• Entry DOI: 10.2210/pdb3vw4/pdb
• Descriptor: Rep, DNA (5'-D(P*AP*AP*TP*GP*AP*GP*AP*CP*CP*AP*GP*AP*TP*AP*AP*GP*CP*CP*TP*TP*AP*TP*C)-3'), DNA (5'-D(P*GP*AP*TP*AP*AP*GP*GP*CP*TP*TP*AP*TP*CP*TP*GP*GP*TP*CP*TP*CP*AP*TP*T)-3'), ... (5 entities in total)
• Functional Keywords: helix-turn-helix, specific dna-binding and unwinding of dna duplex, cytosol, replication initiator protein, dna binding protein-dna complex, dna binding protein/dna
• Biological source: Escherichia coli; More
• Total number of polymer chains: 6
• Total formula weight: 58172.65

Authors

Itou, H.,Yagura, M.,Itoh, T.,Shirakihara, Y. (deposition date: 2012-07-31, release date: 2013-07-31, Last modification date: 2024-03-20)

Primary citation

Itou, H.,Yagura, M.,Shirakihara, Y.,Itoh, T.
Structural Basis for Replication Origin Unwinding by An Initiator-Primase of Plasmid ColE2-P9: Duplex DNA Unwinding by A Single Protein
J.Biol.Chem., 290:3601-3611, 2015

PubMed Abstract: Duplex DNA is generally unwound by protein oligomers prior to replication. The Rep protein of plasmid ColE2-P9 (34 kDa) is an essential initiator for plasmid DNA replication. This protein binds the replication origin (Ori) in a sequence-specific manner as a monomer and unwinds DNA. Here we present the crystal structure of the DNA-binding domain of Rep (E2Rep-DBD) in complex with Ori DNA. The structure unveils the basis for Ori-specific recognition by the E2Rep-DBD and also reveals that it unwinds DNA by the concerted actions of its three contiguous structural modules. The structure also shows that the functionally unknown PriCT domain, which forms a compact module, plays a central role in DNA unwinding. The conservation of the PriCT domain in the C termini of some archaeo-eukaryotic primases indicates that it probably plays a similar role in these proteins. Thus, this is the first report providing the structural basis for the functional importance of the conserved PriCT domain and also reveals a novel mechanism for DNA unwinding by a single protein.

PubMed: 25538245
DOI: 10.1074/jbc.M114.595645

Experimental method

X-RAY DIFFRACTION (2.7 Å)