Nucleic Acids Research Advance Access originally published online on August 9, 2006
Nucleic Acids Research 2006 34(14):3878-3886; doi:10.1093/nar/gkl536
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Nucleic Acids Research, 2006, Vol. 34, No. 14 3878-3886
© 2006 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commerical use, distribution, and reproduction in any medium, provided the original work is properly cited.
Structural Biology |
Complexed crystal structure of replication restart primosome protein PriB reveals a novel single-stranded DNA-binding mode
1 Institute of Molecular Biology, Academia Sinica Taipei, 115, Taiwan 2 Institute of Bioinformatics and Structural Biology, National Tsing Hua University Hsinchu, 300, Taiwan
*To whom correspondence should be addressed. Tel: +886 2 2788 2743; Fax: +886 2 2782 6085; Email: hsiao{at}gate.sinica.edu.tw
Received April 28, 2006. Revised July 4, 2006. Accepted July 13, 2006.
PriB is a primosomal protein required for replication restart in Escherichia coli. PriB stimulates PriA helicase activity via interaction with single-stranded DNA (ssDNA), but the molecular details of this interaction remain unclear. Here, we report the crystal structure of PriB complexed with a 15 bases oligonucleotide (dT15) at 2.7 Å resolution. PriB shares structural similarity with the E.coli ssDNA-binding protein (EcoSSB). However, the structure of the PriB–dT15 complex reveals that PriB binds ssDNA differently. Results from filter-binding assays show that PriB–ssDNA interaction is salt-sensitive and cooperative. Mutational analysis suggests that the loop L45 plays an important role in ssDNA binding. Based on the crystal structure and biochemical analyses, we propose a cooperative mechanism for the binding of PriB to ssDNA and a model for the assembly of the PriA–PriB–ssDNA complex. This report presents the first structure of a replication restart primosomal protein complexed with DNA, and a novel model that explains the interactions between a dimeric oligonucleotide-binding-fold protein and ssDNA.
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
Protein Data Bank accession no. 2ccz
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