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The fundamental problems in duplicating and transmitting genetic information posed by the geometric and topological features of DNA, combined with its large size, are qualitatively similar for prokaryotic and eukaryotic chromosomes. The evolutionary solutions to these problems reveal common themes. However, depending on differences in their organization, ploidy, and copy number, chromosomes and plasmids display distinct segregation strategies as well. In bacteria, chromosome duplication, likely mediated by a stationary replication factory, is accompanied by rapid, directed migration of the daughter duplexes with assistance from DNA-compacting and perhaps translocating proteins. The segregation of unit-copy or low-copy bacterial plasmids is also regulated spatially and temporally by their respective partitioning systems. Eukaryotic chromosomes utilize variations of a basic pairing and unpairing mechanism for faithful segregation during mitosis and meiosis. Rather surprisingly, the yeast plasmid 2-micron circle also resorts to a similar scheme for equal partitioning during mitosis.
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Supplemental Video 1. P1 plasmid segregation. The dynamics of a mini-P1 plasmid in E. coli growing in minimal glucose casamino-acids medium is visualized by the plasmid associated GFP-ParB. Individual frames are taken 5 min. apart (Y. Li and S. Austin, NCI/FCRDC, Frederick, MD)
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Supplemental Video 2. Segregation of the RK2 plasmid. E. coli cells containing RK2 tagged by GFP-LacI/LacO are placed on an agarose slab at 30°C, and images are captured every 30 seconds for five minutes (J. Pogliano, University of California, San Diego, CA). RK2 localizes in clusters tethered to the cell midpoint where they replicate and then separate.
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Supplemental Video 3. Segregation of the 2 micron yeast plasmid. A fluorescence-tagged and 2 micron circle derived reporter plasmid is followed through the cell cycle in a host strain supplying the partitioning proteins (S. Velmurugan and M. Jayaram, University of Texas, Austin, TX). Photographs are taken at 6 min intervals over a period of 90 min.
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Supplemental Video 4. Chromosome segregation in C. crescentus. Swarmer cells in which the chromosomal origin is fluorescence-tagged (LacI-ecfp/LacO) are transferred onto a nutrient-containing agarose pad. Images are taken at 2 min intervals to follow the duplication and segregation of the origin region (P. H. Viollier, M. Thanbichler, L. Shapiro and colleagues, Stanford University, Stanford, CA).
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Supplemental Video 5. Oscillations of the bacterial cytoskeletal ring protein FtsZ. In this time lapse movie of immobilized E. coli cells expressing FtsZ-GFP, The Z rings are visible, as are the pole to pole oscillations. A spiral pattern is sometimes discernible in these oscillations (provided by W. Margolin at the University of Houston. TX).
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Supplemental Video 6. Oscillations of the MinD protein. This time lapse movie (courtesy of W. Margolin, University of Houston, TX) depicts several oscillation cycles of the MinD protein in E. coli cells expressing GFP-MinD as well as MinE (untagged).
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