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First published online August 24, 2006
doi: 10.1242/10.1242/jcs.03066

Journal of Cell Science 119, 3571-3582 (2006)
Published by The Company of Biologists 2006
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Research Article

Decreased polarity and increased random motility in PtK1 epithelial cells correlate with inhibition of endosomal recycling

Natalie L. Prigozhina1 and Clare M. Waterman-Storer2

1 The Burnham Institute, 10901 N. Torrey Pines Road, Room 7108, La Jolla, CA 92037, USA
2 The Scripps Research Institute, Department of Cell Biology, CB163, 10550 N. Torrey Pines Road, La Jolla, CA 92037, USA

Authors for correspondence (e-mail: nataliep{at}burnham.org; waterman{at}scripps.edu)

Accepted 24 May 2006

Locomoting cells exhibit a constant retrograde flow of plasma membrane proteins from the leading edge towards the cell center, which, when coupled to substrate adhesion, may drive forward cell movement. Here, we aimed to test the hypothesis that, in epithelial cells, these plasma membrane components are delivered via a polarized endo/exocytotic cycle, and that their correct recycling is required for normal migration. To this end, we expressed in PtK1 cells cDNA constructs encoding GDP-restricted (S25N) and GTP-restricted (Q70L) mutants of Rab11b, a small GTPase that has been implicated in the late stage of recycling, where membrane components from the endosomal recycling compartment are transported back to the plasma membrane. Surprisingly, we found that transient expression of the Rab11b mutants in randomly migrating PtK1 cells in small cell islands caused altered cell morphology and actually increased the velocity of cell locomotion. Stable expression of either mutant protein also did not decrease cell migration velocity, but instead affected the directionality of migration in monolayer wound healing assays. We have also tested the effects of other Rab proteins, implicated in endocytic recycling, and discovered a clear correlation between the degree of recycling inhibition and the increase in non-directional cell motility.

Key words: Cell motility, Recycling, Directional migration, Membrane traffic, Epithelial cells, Cell morphology


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