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J Biol Chem. 2012 Jul 27;287(31):26146-54. doi: 10.1074/jbc.M112.373696. Epub 2012 Jun 6.

Structural mechanism of ATP-induced polymerization of the partition factor ParF: implications for DNA segregation.

Author information

1
Department of Biochemistry, Duke University School of Medicine, Durham, North Carolina 27710, USA. maria.schumacher@duke.edu

Abstract

Segregation of the bacterial multidrug resistance plasmid TP228 requires the centromere-binding protein ParG, the parH centromere, and the Walker box ATPase ParF. The cycling of ParF between ADP- and ATP-bound states drives TP228 partition; ATP binding stimulates ParF polymerization, which is essential for segregation, whereas ADP binding antagonizes polymerization and inhibits DNA partition. The molecular mechanism involved in this adenine nucleotide switch is unclear. Moreover, it is unknown how any Walker box protein polymerizes in an ATP-dependent manner. Here, we describe multiple ParF structures in ADP- and phosphomethylphosphonic acid adenylate ester (AMPPCP)-bound states. ParF-ADP is monomeric but dimerizes when complexed with AMPPCP. Strikingly, in ParF-AMPPCP structures, the dimers interact to create dimer-of-dimer "units" that generate a specific linear filament. Mutation of interface residues prevents both polymerization and DNA segregation in vivo. Thus, these data provide insight into a unique mechanism by which a Walker box protein forms polymers that involves the generation of ATP-induced dimer-of-dimer building blocks.

PMID:
22674577
PMCID:
PMC3406698
DOI:
10.1074/jbc.M112.373696
[Indexed for MEDLINE]
Free PMC Article

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