Nucleic Acids Research Advance Access originally published online on May 13, 2008
Nucleic Acids Research 2008 36(11):3676-3689; doi:10.1093/nar/gkn170
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Nucleic Acids Research, 2008, Vol. 36, No. 11 3676-3689
© 2008 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-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Molecular Biology |
Streptococcus pyogenes pSM19035 requires dynamic assembly of ATP-bound ParA and ParB on parS DNA during plasmid segregation
1Department of Microbial Biotechnology, National Centre of Biotechnology, CSIC, 28049 Madrid, Spain, 2Institute of Chemistry and Biochemistry / Crystallography, Freie Universität Berlin, 14195 and 3Max-Planck Institute for Molecular Genetics, 14195 Berlin, Germany
*To whom correspondence should be addressed. Tel: +34 91585 4546; Fax: +34 91585 4506; Email: jcalonso{at}cnb.csic.es
Received January 8, 2008. Revised March 19, 2008. Accepted March 25, 2008.
The accurate partitioning of Firmicute plasmid pSM19035 at cell division depends on ATP binding and hydrolysis by homodimeric ATPase 
2 (ParA) and binding of 
2 (ParB) to its cognate parS DNA. The 1.83 Å resolution crystal structure of 2 in a complex with non-hydrolyzable ATP
S reveals a unique ParA dimer assembly that permits nucleotide exchange without requiring dissociation into monomers. In vitro, 2 had minimal ATPase activity in the absence of 2 and parS DNA. However, stoichiometric amounts of 2 and parS DNA stimulated the 2 ATPase activity and mediated plasmid pairing, whereas at high (4:1) 2 : 2 ratios, stimulation of the ATPase activity was reduced and 2 polymerized onto DNA. Stimulation of the 2 ATPase activity and its polymerization on DNA required ability of 2 to bind parS DNA and its N-terminus. In vivo experiments showed that 2 alone associated with the nucleoid, and in the presence of 2 and parS DNA, 2 oscillated between the nucleoid and the cell poles and formed spiral-like structures. Our studies indicate that the molar 2 : 2 ratio regulates the polymerization properties of (•ATP•Mg2+)2 on and depolymerization from parS DNA, thereby controlling the temporal and spatial segregation of pSM19035 before cell division.
Present address: Wilhelm A. Weihofen, Department of Molecular and Cellular Biology, Harvard University, MA 02138, USA
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
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