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          Institute: MPI für molekulare Genetik     Collection: Microscopy     Display Documents



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ID: 412078.0, MPI für molekulare Genetik / Microscopy
Streptococcus pyogenes pSM19035 requires dynamic assembly of ATP-bound ParA and ParB on parS DNA during plasmid segregation
Authors:Pratto, Florencia; Cicek, Aslan; Weihofen, Wilhelm A.; Lurz, Rudi; Saenger, Wolfram; Alonso, Juan C.
Language:English
Date of Publication (YYYY-MM-DD):2008-05-13
Title of Journal:Nucleic Acids Research
Journal Abbrev.:Nucleic Acids Res.
Volume:36
Issue / Number:11
Start Page:3676
End Page:3689
Copyright:© 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.
Review Status:not specified
Audience:Experts Only
Abstract / Description:The accurate partitioning of Firmicute plasmid pSM19035 at cell division depends on ATP binding and hydrolysis by homodimeric ATPase {delta}2 (ParA) and binding of {omega}2 (ParB) to its cognate parS DNA. The 1.83 Å resolution crystal structure of {delta}2 in a complex with non-hydrolyzable ATP{gamma}S reveals a unique ParA dimer assembly that permits nucleotide exchange without requiring dissociation into monomers. In vitro, {delta}2 had minimal ATPase activity in the absence of {omega}2 and parS DNA. However, stoichiometric amounts of {omega}2 and parS DNA stimulated the {delta}2 ATPase activity and mediated plasmid pairing, whereas at high (4:1) {omega}2 : {delta}2 ratios, stimulation of the ATPase activity was reduced and {delta}2 polymerized onto DNA. Stimulation of the {delta}2 ATPase activity and its polymerization on DNA required ability of {omega}2 to bind parS DNA and its N-terminus. In vivo experiments showed that {delta}2 alone associated with the nucleoid, and in the presence of {omega}2 and parS DNA, {delta}2 oscillated between the nucleoid and the cell poles and formed spiral-like structures. Our studies indicate that the molar {omega}2 : {delta}2 ratio regulates the polymerization properties of ({delta}•ATP•Mg2+)2 on and depolymerization from parS DNA, thereby controlling the temporal and spatial segregation of pSM19035 before cell division.
Comment of the Author/Creator:To whom correspondence should be addressed. Tel: +34 91585 4546; Fax: +34 91585 4506; Email: jcalonso@cnb.csic.es
External Publication Status:published
Document Type:Article
Communicated by:Rudi Lurz
Affiliations:MPI für molekulare Genetik
External Affiliations:1.Department of Microbial Biotechnology, National Centre of Biotechnology, CSIC, 28049 Madrid, Spain;
2.Institute of Chemistry and Biochemistry / Crystallography, Freie Universität Berlin, Germany.
Identifiers:URL:http://nar.oxfordjournals.org/cgi/reprint/36/11/36...
DOI:10.1093/nar/gkn170
ISSN:0305-1048
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