A novel high-content analysis tool reveals Rab8-driven cytoskeletal reorganization through Rho GTPases, calpain and MT1-MMP

Bravo‐Cordero, Jose Javier; Cordani, Marco; Soriano, Silvia F.; Díez, Begoña; Muñoz-Agudo, Carmen; Casanova-Acebes, María; Boullosa, César; Guadamillas, Marta C.; Ezkurdia, Iakes; González-Pisano, David; Pozo, Miguel Á. del; Montoya, Marı́a C.

doi:10.1242/jcs.174920

Cited by 26 publications

(35 citation statements)

References 61 publications

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“…In agreement with earlier studies, we found that in HT 1080 cells grown on collagen-coated dishes, export of MMP14 was inhibited by overexpression of Rab8a T22N ( Fig. 8 A; Bravo-Cordero et al, 2016Wiesner et al, 2013). It was also inhibited by Rab10 T23N, MICAL1 G3W, GRAF1/2 BAR-PH, and WDR44 ΔC.…”

Section: Mical1 Connects Graf1b/2 To Rab8 and Rab10supporting

confidence: 92%

GRAF2, WDR44, and MICAL1 mediate Rab8/10/11–dependent export of E-cadherin, MMP14, and CFTR ΔF508

Häsler

Vallis

Pasche

et al. 2020

Journal of Cell Biology

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In addition to the classical pathway of secretion, some transmembrane proteins reach the plasma membrane through alternative routes. Several proteins transit through endosomes and are exported in a Rab8-, Rab10-, and/or Rab11-dependent manner. GRAFs are membrane-binding proteins associated with tubules and vesicles. We found extensive colocalization of GRAF1b/2 with Rab8a/b and partial with Rab10. We identified MICAL1 and WDR44 as direct GRAF-binding partners. MICAL1 links GRAF1b/2 to Rab8a/b and Rab10, and WDR44 binds Rab11. Endogenous WDR44 labels a subset of tubular endosomes, which are closely aligned with the ER via binding to VAPA/B. With its BAR domain, GRAF2 can tubulate membranes, and in its absence WDR44 tubules are not observed. We show that GRAF2 and WDR44 are essential for the export of neosynthesized E-cadherin, MMP14, and CFTR ΔF508, three proteins whose exocytosis is sensitive to ER stress. Overexpression of dominant negative mutants of GRAF1/2, WDR44, and MICAL1 also interferes with it, facilitating future studies of Rab8/10/11–dependent exocytic pathways of central importance in biology.

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Section: Mical1 Connects Graf1b/2 To Rab8 and Rab10supporting

confidence: 92%

GRAF2, WDR44, and MICAL1 mediate Rab8/10/11–dependent export of E-cadherin, MMP14, and CFTR ΔF508

Häsler

Vallis

Pasche

et al. 2020

Journal of Cell Biology

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“…57 Most recently, Rab8 activation has been shown to induce Rac1 activity, causing cortical actin formation, focal adhesion disassembly, and stress fiber disassembly. 58 This effect was dependent on Calpain, MT1-MMP, and Rho GTPases. 58 Furthermore, Rab8 is necessary for persistent cell motility, as well as EGFinduced polarity and chemotaxis.…”

Section: Molecular Control Of Calcium Influxes Upon Induction Of Epitmentioning

confidence: 97%

“…58 Furthermore, Rab8 is necessary for persistent cell motility, as well as EGFinduced polarity and chemotaxis. 58 These data all point to a direct link between the slow endocytic recycling pathway and initiation of cellular motility required for EMT. Additionally, it shows Rab8 affects RhoA activity.…”

Section: Molecular Control Of Calcium Influxes Upon Induction Of Epitmentioning

confidence: 99%

Control of cell mechanics by RhoA and calcium fluxes during epithelial scattering

2016

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Epithelial tissues use adherens junctions to maintain tight interactions and coordinate cellular activities. Adherens junctions are remodeled during epithelial morphogenesis, including instances of epithelialmesenchymal transition, or EMT, wherein individual cells detach from the tissue and migrate as individual cells. EMT has been recapitulated by growth factor induction of epithelial scattering in cell culture. In culture systems, cells undergo a highly reproducible series of cell morphology changes, most notably cell spreading followed by cellular compaction and cell migration. These morphology changes are accompanied by striking actin rearrangements. The current evidence suggests that global changes in actomyosin-based cellular contractility, first a loss of contractility during spreading and its activation during cell compaction, are the main drivers of epithelial scattering. In this review, we focus on how spreading and contractility might be controlled during epithelial scattering. While we propose a central role for RhoA, which is well known to control cellular contractility in multiple systems and whose role in epithelial scattering is well accepted, we suggest potential roles for additional cellular systems whose role in epithelial cell biology has been less well documented. In particular, we propose critical roles for vesicle recycling, calcium channels, and calcium-dependent kinases.

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“…In fact, FIP3 silencing induced T-cell spreading, a process that is controlled by Rac1, suggesting endosomal trafficking of Rac1 to regulate T-cell spreading and activation in the immunological synapse [131]. Finally, Rab8, which localizes to the endocytic recycling compartment, was demonstrated to increase Rac1 activity and Tiam1/Rac1 mobilization from intracellular compartments to cortical locations to maintain directionality of migrating cells by enabling focal adhesion turnover and actin polymerization [132]. Combined, these studies suggest that endosomal trafficking routes are major determinants of Rac activity patterns emanating from the PM.…”

Section: Rho Gtpases Acting Along the Endocytic Pathwaymentioning

confidence: 96%

Spatiotemporal Control of Intracellular Membrane Trafficking by Rho GTPases

Noll

Haußer

2019

Cells

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As membrane-associated master regulators of cytoskeletal remodeling, Rho GTPases coordinate a wide range of biological processes such as cell adhesion, motility, and polarity. In the last years, Rho GTPases have also been recognized to control intracellular membrane sorting and trafficking steps directly; however, how Rho GTPase signaling is regulated at endomembranes is still poorly understood. In this review, we will specifically address the local Rho GTPase pools coordinating intracellular membrane trafficking with a focus on the endo-and exocytic pathways. We will further highlight the spatiotemporal molecular regulation of Rho signaling at endomembrane sites through Rho regulatory proteins, the GEFs and GAPs. Finally, we will discuss the contribution of dysregulated Rho signaling emanating from endomembranes to the development and progression of cancer.an inactive GDP-bound and an active GTP-bound state. Rho GDP/GTP cycling is tightly regulated by guanine nucleotide exchange factors (GEFs) that promote the formation of the active GTP-bound form through exchanging GDP for GTP. On the contrary, GTPase-activating proteins (GAPs) catalyze the intrinsic GTPase activity and promote the formation of inactive GDP-bound Rho. This inactive state can be subsequently recognized by guanine nucleotide dissociation inhibitors (GDIs), which sequester Rho GTPases in the cytosol [5,6] (Figure 1). Whereas classical Rho GTPases are regulated by GDP/GTP cycling, atypical Rho GTPases are regulated by other mechanisms that occur in particular at the transcriptional and post-translational level [7]. In the case of atypical Rho GTPases, GTP is usually constitutively bound, either because these Rho GTPases possess high intrinsic nucleotide exchange activity or have substitutions in their GTPase domain that prevent GTP hydrolysis.

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A novel high-content analysis tool reveals Rab8-driven cytoskeletal reorganization through Rho GTPases, calpain and MT1-MMP

Cited by 26 publications

References 61 publications

GRAF2, WDR44, and MICAL1 mediate Rab8/10/11–dependent export of E-cadherin, MMP14, and CFTR ΔF508

GRAF2, WDR44, and MICAL1 mediate Rab8/10/11–dependent export of E-cadherin, MMP14, and CFTR ΔF508

Control of cell mechanics by RhoA and calcium fluxes during epithelial scattering

Spatiotemporal Control of Intracellular Membrane Trafficking by Rho GTPases

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