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Stress fiber-based movement of Shigella flexneri within cells
T Vasselon1, J Mounier, M C Prevost
1Unité de Pathogénie Microbienne Moléculaire, Institut Pasteur, Paris, France.
Abstract:
icsA (virG), a gene located on pWR100, the virulence plasmid of Shigella flexneri serotype 5 (M90T), encodes a 120-kDa outer membrane protein. This protein promotes a random intracellular movement of the bacteria and leads to the infection of adjacent cells by the formation of protrusions. This movement, which involves the nucleation, polymerization, and subsequent polarization of actin, is referred to as the Ics phenotype (intra/intercellular spread). Here we present evidence that a second, distinct form of locomotion is also elaborated by S. flexneri in chicken embryo fibroblasts in which the Ics phenotype is not expressed. Using a combination of phase-contrast microcinematography and confocal microscopy, we have demonstrated that bacteria adopt parallel orientations by interacting with stress fibers. This interaction subsequently results in bacterial movement along the stress fibers themselves. This phenomenon occurs independently of the presence of a phagocytic vacuole which is lysed shortly after entry of the bacteria into the cell. It is expressed by M90T and SC560, its icsA mutant. This movement has been termed organelle-like movement (Olm phenotype) and is thought to account for the early accumulation of bacteria seen near the nucleus.
Insights
Shigella flexneri exhibits two distinct movement types within host cells. The known IcsA-dependent intracellular spread (Ics phenotype) and a novel actin-independent organelle-like movement (Olm phenotype) along stress fibers.
Area of Science:
- Microbiology
- Cell Biology
- Bacterial Pathogenesis
Background:
- Shigella flexneri uses the IcsA protein for intracellular movement and cell-to-cell spread (Ics phenotype).
- This process involves actin polymerization and bacterial protrusions.
Purpose of the Study:
- To investigate alternative bacterial locomotion mechanisms in S. flexneri.
- To characterize a novel movement phenotype independent of the IcsA protein.
Main Methods:
- Phase-contrast microcinematography
- Confocal microscopy
- Analysis of S. flexneri M90T and its icsA mutant (SC560) in chicken embryo fibroblasts.
Main Results:
- A second form of locomotion, the organelle-like movement (Olm phenotype), was observed.
- Olm phenotype involves bacteria aligning with and moving along cellular stress fibers.
- This movement is independent of the IcsA protein and phagocytic vacuole lysis.
Conclusions:
- S. flexneri possesses at least two distinct motility mechanisms within host cells.
- The Olm phenotype may explain early bacterial aggregation near the host cell nucleus.