Inhibition of protein and RNA synthesis in Escherichia coli results in declustering of plasmid RK2

Multi-copy plasmids in Escherichia coli are not randomly distributed throughout the cell but are present as clusters of plasmid molecules that are localized at preferred cellular locations. A plasmid RK2 derivative (pZZ15) that can be tagged with a green fluorescent protein-LacI fusion protein normally exists as clusters that are localized at the mid- and quarter-cell positions. In this study the effect of the protein synthesis inhibitor, chloramphenicol, and the RNA synthesis inhibitor, rifampicin, on RK2 clustering and localization was examined. The addition of either inhibitor to exponentially growing E. coli cells carrying pZZ15 results in a displacement of the position and a declustering of this multi-copy plasmid indicating that continued protein synthesis and RNA synthesis are required for clustering and localization of this plasmid. It is likely that it is not just the process of transcription or translation that is important for clustering but rather some host or plasmid encoded factor(s) that is required.