2NCJ
Solution Structure of the PriC DNA replication restart protein
Summary for 2NCJ
| Entry DOI | 10.2210/pdb2ncj/pdb |
| NMR Information | BMRB: 26026 |
| Descriptor | Uncharacterized protein (1 entity in total) |
| Functional Keywords | dna replication restart, dna binding protein |
| Biological source | Cronobacter sakazakii ATCC BAA-894 |
| Total number of polymer chains | 1 |
| Total formula weight | 19733.47 |
| Authors | Cornilescu, C.C.,Cornilescu, G.,Wessel, S.R.,Keck, J.L.,Markley, J.L. (deposition date: 2016-04-07, release date: 2016-07-06, Last modification date: 2024-05-15) |
| Primary citation | Wessel, S.R.,Cornilescu, C.C.,Cornilescu, G.,Metz, A.,Leroux, M.,Hu, K.,Sandler, S.J.,Markley, J.L.,Keck, J.L. Structure and Function of the PriC DNA Replication Restart Protein. J.Biol.Chem., 291:18384-18396, 2016 Cited by PubMed Abstract: Collisions between DNA replication complexes (replisomes) and barriers such as damaged DNA or tightly bound protein complexes can dissociate replisomes from chromosomes prematurely. Replisomes must be reloaded under these circumstances to avoid incomplete replication and cell death. Bacteria have evolved multiple pathways that initiate DNA replication restart by recognizing and remodeling abandoned replication forks and reloading the replicative helicase. In vitro, the simplest of these pathways is mediated by the single-domain PriC protein, which, along with the DnaC helicase loader, can load the DnaB replicative helicase onto DNA bound by the single-stranded DNA (ssDNA)-binding protein (SSB). Previous biochemical studies have identified PriC residues that mediate interactions with ssDNA and SSB. However, the mechanisms by which PriC drives DNA replication restart have remained poorly defined due to the limited structural information available for PriC. Here, we report the NMR structure of full-length PriC from Cronobacter sakazakii PriC forms a compact bundle of α-helices that brings together residues involved in ssDNA and SSB binding at adjacent sites on the protein surface. Disruption of these interaction sites and of other conserved residues leads to decreased DnaB helicase loading onto SSB-bound DNA. We also demonstrate that PriC can directly interact with DnaB and the DnaB·DnaC complex. These data lead to a model in which PriC acts as a scaffold for recruiting DnaB·DnaC to SSB/ssDNA sites present at stalled replication forks. PubMed: 27382050DOI: 10.1074/jbc.M116.738781 PDB entries with the same primary citation |
| Experimental method | SOLUTION NMR |
Structure validation
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