Dynamic movement of actin-like proteins within bacterial cells

Abstract

Actin proteins are present in pro- and eukaryotes, and have been shown to perform motor-like functions in eukaryotic cells in a variety of processes. Bacterial actin homologues are essential for cell viability and have been implicated in the formation of rod cell shape, as well as in segregation of plasmids and whole chromosomes. We have generated functional green fluorescent protein fusions of all three Bacillus subtilis actin-like proteins (MreB, MbI and MreBH), and show that all three proteins form helical filaments underneath the cell membrane, the pattern of which is distinct for each protein. Time-lapse microscopy showed that the filaments are highly dynamic structures. A number of separate filaments of MreB and MbI continuously move through the cell along helical tracks underneath the cell membrane. The speed of extension of the growing end of filaments is within the range of known actin polymerization (0.1 μm/s), generating a potential poleward or centreward pushing velocity at 0.24μm/min for MreB or MbI, respectively. During nutritional downshift and a block in topoisomerase IV activity, the filaments rapidly disintegrated, showing that movement occurs only in growing cells. Contrary to MbI and MreBH filaments, MreB filaments were generally absent in cells lacking DNA, providing a further distinction between the three orthologues. © 2004 EUROPEAN MOLECULAR BIOLOGY ORGANIZATION.