4OU7

Crystal structure of DnaT84-153-dT10 ssDNA complex reveals a novel single-stranded DNA binding mode

Summary for 4OU7

• Entry DOI: 10.2210/pdb4ou7/pdb
• Related: 4OU6
• Descriptor: Primosomal protein 1, DNA (5'-D(P*TP*TP*TP*TP*TP*TP*TP*TP*TP*T)-3') (2 entities in total)
• Functional Keywords: dna binding, replication-dna complex, replication/dna
• Biological source: Escherichia coli
• Total number of polymer chains: 6
• Total formula weight: 43838.70

Authors

Liu, Z.,Chen, P.,Niu, L.,Teng, M.,Li, X. (deposition date: 2014-02-15, release date: 2014-08-13, Last modification date: 2024-05-29)

Primary citation

Liu, Z.,Chen, P.,Wang, X.,Cai, G.,Niu, L.,Teng, M.,Li, X.
Crystal structure of DnaT84-153-dT10 ssDNA complex reveals a novel single-stranded DNA binding mode.
Nucleic Acids Res., 42:9470-9483, 2014

PubMed Abstract: DnaT is a primosomal protein that is required for the stalled replication fork restart in Escherichia coli. As an adapter, DnaT mediates the PriA-PriB-ssDNA ternary complex and the DnaB/C complex. However, the fundamental function of DnaT during PriA-dependent primosome assembly is still a black box. Here, we report the 2.83 Å DnaT(84-153)-dT10 ssDNA complex structure, which reveals a novel three-helix bundle single-stranded DNA binding mode. Based on binding assays and negative-staining electron microscopy results, we found that DnaT can bind to phiX 174 ssDNA to form nucleoprotein filaments for the first time, which indicates that DnaT might function as a scaffold protein during the PriA-dependent primosome assembly. In combination with biochemical analysis, we propose a cooperative mechanism for the binding of DnaT to ssDNA and a possible model for the assembly of PriA-PriB-ssDNA-DnaT complex that sheds light on the function of DnaT during the primosome assembly and stalled replication fork restart. This report presents the first structure of the DnaT C-terminal complex with ssDNA and a novel model that explains the interactions between the three-helix bundle and ssDNA.

PubMed: 25053836
DOI: 10.1093/nar/gku633

Experimental method

X-RAY DIFFRACTION (2.83 Å)