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, 52 (2), 385-98

Bacterial Mitosis: Partitioning Protein ParA Oscillates in Spiral-Shaped Structures and Positions Plasmids at Mid-Cell

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Bacterial Mitosis: Partitioning Protein ParA Oscillates in Spiral-Shaped Structures and Positions Plasmids at Mid-Cell

Gitte Ebersbach et al. Mol Microbiol.

Abstract

The par2 locus of Escherichia coli plasmid pB171 encodes oscillating ATPase ParA, DNA binding protein ParB and two cis-acting DNA regions to which ParB binds (parC1 and parC2). Three independent techniques were used to investigate the subcellular localization of plasmids carrying par2. In cells with a single plasmid focus, the focus located preferentially at mid-cell. In cells with two foci, these located at quarter-cell positions. In the absence of ParB and parC1/parC2, ParA-GFP formed stationary helices extending from one end of the nucleoid to the other. In the presence of ParB and parC1/parC2, ParA-GFP oscillated in spiral-shaped structures. Amino acid substitutions in ParA simultaneously abolished ParA spiral formation, oscillation and either plasmid localization or plasmid separation at mid-cell. Therefore, our results suggest that ParA spirals position plasmids at the middle of the bacterial nucleoid and subsequently separate them into daughter cells.

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