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MBC in Press, published online ahead of print September 29, 2004
Mol. Biol. Cell 10.1091/mbc.E04-06-0509

A more recent version of this article appeared on December 1, 2004
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Submitted on June 22, 2004
Revised on September 17, 2004
Accepted on September 20, 2004

Repeated Cycles of Rapid Actin Assembly and Disassembly on Epithelial Cell Phagosomes

Patricia T. Yam* and Julie A. Theriot*{dagger}{ddagger}

Departments of *Biochemistry and {dagger}Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305

Monitoring Editor: Jennifer Lippincott-Schwartz

We have found that early in infection of the intracellular pathogen Listeria monocytogenes in MDCK epithelial cells expressing GFP-actin, F-actin rapidly assembles (~25 s) and disassembles (~30 s) around the bacteria - a phenomenon we call flashing. L. monocytogenes strains unable to perform actin-based motility or unable to escape the phagosome were capable of flashing, suggesting that the actin assembly occurs on the phagosome membrane. Cycles of actin assembly and disassembly could occur repeatedly on the same phagosome. Indirect immunofluorescence showed that most bacteria were fully internalized when flashing occurred, suggesting that actin flashing does not represent phagocytosis. E. coli expressing invA, a gene product from Yersinia pseudotuberculosis that mediates cellular invasion, also induced flashing. Furthermore, polystyrene beads coated with E-cadherin or transferrin also induced flashing after internalization. This suggests that flashing occurs downstream of several distinct molecular entry mechanisms and may be a general consequence of internalization of large objects by epithelial cells.

{ddagger}Corresponding author. E-mail: theriot{at}cmgm.stanford.edu






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