ParA2, a Vibrio cholerae chromosome partitioning protein, forms left-handed helical filaments on DNA.

Details

Serval ID
serval:BIB_6FAEA31F23C9
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
ParA2, a Vibrio cholerae chromosome partitioning protein, forms left-handed helical filaments on DNA.
Journal
Proceedings of the National Academy of Sciences of the United States of America
Author(s)
Hui M.P., Galkin V.E., Yu X., Stasiak A.Z., Stasiak A., Waldor M.K., Egelman E.H.
ISSN
1091-6490[electronic], 0027-8424[linking]
Publication state
Published
Issued date
2010
Volume
107
Number
10
Pages
4590-4595
Language
english
Abstract
Most bacterial chromosomes contain homologs of plasmid partitioning (par) loci. These loci encode ATPases called ParA that are thought to contribute to the mechanical force required for chromosome and plasmid segregation. In Vibrio cholerae, the chromosome II (chrII) par locus is essential for chrII segregation. Here, we found that purified ParA2 had ATPase activities comparable to other ParA homologs, but, unlike many other ParA homologs, did not form high molecular weight complexes in the presence of ATP alone. Instead, formation of high molecular weight ParA2 polymers required DNA. Electron microscopy and three-dimensional reconstruction revealed that ParA2 formed bipolar helical filaments on double-stranded DNA in a sequence-independent manner. These filaments had a distinct change in pitch when ParA2 was polymerized in the presence of ATP versus in the absence of a nucleotide cofactor. Fitting a crystal structure of a ParA protein into our filament reconstruction showed how a dimer of ParA2 binds the DNA. The filaments formed with ATP are left-handed, but surprisingly these filaments exert no topological changes on the right-handed B-DNA to which they are bound. The stoichiometry of binding is one dimer for every eight base pairs, and this determines the geometry of the ParA2 filaments with 4.4 dimers per 120 A pitch left-handed turn. Our findings will be critical for understanding how ParA proteins function in plasmid and chromosome segregation.
Keywords
Adenosine Diphosphate/chemistry, Adenosine Diphosphate/metabolism, Adenosine Triphosphatases/chemistry, Adenosine Triphosphatases/metabolism, Adenosine Triphosphate/chemistry, Adenosine Triphosphate/metabolism, Bacterial Proteins/chemistry, Bacterial Proteins/metabolism, Chromosomes, Bacterial/genetics, DNA/chemistry, DNA/metabolism, DNA-Binding Proteins/chemistry, DNA-Binding Proteins/metabolism, Hydrolysis, Kinetics, Microscopy, Electron, Models, Molecular, Plasmids/genetics, Protein Binding, Protein Multimerization, Protein Structure, Secondary, Protein Structure, Tertiary, Vibrio cholerae/genetics, Vibrio cholerae/metabolism
Pubmed
Web of science
Open Access
Yes
Create date
26/11/2010 12:27
Last modification date
20/08/2019 15:28
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