Rab14 and Its Exchange Factor FAM116 Link Endocytic Recycling and Adherens Junction Stability in Migrating Cells

Andrea Linford,Shin-Ichiro Yoshimura,Ricardo Nunes Bastos,Lars Langemeyer,Andreas Gerondopoulos,Daniel J Rigden,Francis A Barr

doi:10.1016/j.devcel.2012.04.010

Andrea Linford, Shin-Ichiro Yoshimura + Show 5 more

Open Access

https://doi.org/10.1016/j.devcel.2012.04.010

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Abstract

SummaryRab GTPases define the vesicle trafficking pathways underpinning cell polarization and migration. Here, we find that Rab4, Rab11, and Rab14 and the candidate Rab GDP-GTP exchange factors (GEFs) FAM116A and AVL9 are required for cell migration. Rab14 and its GEF FAM116A localize to and act on an intermediate compartment of the transferrin-recycling pathway prior to Rab11 and after Rab5 and Rab4. This Rab14 intermediate recycling compartment has specific functions in migrating cells discrete from early and recycling endosomes. Rab14-depleted cells show increased N-cadherin levels at junctional complexes and cannot resolve cell-cell junctions. This is due to decreased shedding of cell-surface N-cadherin by the ADAM family protease ADAM10/Kuzbanian. In FAM116A- and Rab14-depleted cells, ADAM10 accumulates in a transferrin-positive endocytic compartment, and the cell-surface level of ADAM10 is correspondingly reduced. FAM116 and Rab14 therefore define an endocytic recycling pathway needed for ADAM protease trafficking and regulation of cell-cell junctions.

Highlights

Cell migration and polarization are underpinned by a complex network of cellular trafficking pathways, which transport a variety of different membrane proteins required for sensing extracellular cues, as well as the creation and remodeling of cell adhesions and cell-cell junctions (Baum and Georgiou, 2011; Caswell et al, 2009; Huttenlocher, 2005; Mellman and Nelson, 2008; Ulrich and Heisenberg, 2009)
To identify trafficking pathways required for cell migration in vitro, this scratch-wound cell migration assay was combined with siRNA depletion of human Rab GTPases (Figure 1)
Consistent with previous studies showing that Rab4 and Rab11 are required for integrin and E-cadherin trafficking, Rab4- and Rab11-depleted cells migrated more slowly away from the edge of the cell sheet and failed to close the wound after 16 hr (Figure 1B)

Summary

Introduction

Cell migration and polarization are underpinned by a complex network of cellular trafficking pathways, which transport a variety of different membrane proteins required for sensing extracellular cues, as well as the creation and remodeling of cell adhesions and cell-cell junctions (Baum and Georgiou, 2011; Caswell et al, 2009; Huttenlocher, 2005; Mellman and Nelson, 2008; Ulrich and Heisenberg, 2009). Rab family GTPases are typically associated with different stages of the endocytic recycling pathway (de Renzis et al, 2002; Sonnichsen et al, 2000; Ullrich et al, 1996), for which the transferrin receptor is the most studied cargo

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Discussion

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