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Bipolar elongation of filaments of the bacterial actin homolog ParM drives movement of newly replicated plasmid DNA to opposite poles of a bacterial cell. We used a combination of vitreous sectioning and electron cryotomography to study this DNA partitioning system directly in native, frozen cells. The diffraction patterns from overexpressed ParM bundles in electron cryotomographic reconstructions were used to unambiguously identify ParM filaments in Escherichia coli cells. Using a low-copy number plasmid encoding components required for partitioning, we observed small bundles of three to five intracellular ParM filaments that were situated close to the edge of the nucleoid. We propose that this may indicate the capture of plasmid DNA within the periphery of this loosely defined, chromosome-containing region.

Original publication

DOI

10.1126/science.1164346

Type

Journal article

Journal

Science

Publication Date

23/01/2009

Volume

323

Pages

509 - 512

Keywords

Actins, Cryoelectron Microscopy, Cytoskeleton, DNA, Bacterial, Escherichia coli, Escherichia coli Proteins, Image Processing, Computer-Assisted, Imaging, Three-Dimensional, Plasmids