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Cell Microbiol. 2013 Mar;15(3):353-367. doi: 10.1111/cmi.12051. Epub 2012 Nov 13.

Myosin-X facilitates Shigella-induced membrane protrusions and cell-to-cell spread.

Author information

1
Department of Medicine, Division of Infectious Diseases, University of Florida College of Medicine, Gainesville, FL, USA.
2
Department of Cell and Molecular Physiology, University of North Carolina, Chapel Hill, NC, USA.
3
Department of Medicine, Division of Translational Medicine, Brigham and Women's Hospital, Harvard University, Boston, MA, USA.
#
Contributed equally

Abstract

The intracellular pathogen Shigella flexneri forms membrane protrusions to spread from cell to cell. As protrusions form, myosin-X (Myo10) localizes to Shigella. Electron micrographs of immunogold-labelled Shigella-infected HeLa cells reveal that Myo10 concentrates at the bases and along the sides of bacteria within membrane protrusions. Time-lapse video microscopy shows that a full-length Myo10 GFP-construct cycles along the sides of Shigella within the membrane protrusions as these structures progressively lengthen. RNAi knock-down of Myo10 is associated with shorter protrusions with thicker stalks, and causes a >80% decrease in confluent cell plaque formation. Myo10 also concentrates in membrane protrusions formed by another intracellular bacteria, Listeria, and knock-down of Myo10 also impairs Listeria plaque formation. In Cos7 cells (contain low concentrations of Myo10), the expression of full-length Myo10 nearly doubles Shigella-induced protrusion length, and lengthening requires the head domain, as well as the tail-PH domain, but not the FERM domain. The GFP-Myo10-HMM domain localizes to the sides of Shigella within membrane protrusions and the GFP-Myo10-PH domain localizes to host cell membranes. We conclude thatMyo10 generates the force to enhance bacterial-induced protrusions by binding its head region to actin filaments and its PH tail domain to the peripheral membrane.

PMID:
23083060
PMCID:
PMC4070382
DOI:
10.1111/cmi.12051
[Indexed for MEDLINE]
Free PMC Article

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