Involvement of the Arp2/3 Complex and Scar2 in Golgi Polarity in Scratch Wound Models

Magdalena, Juana; Millard, Thomas H.; Etienne-Manneville, Sandrine; Launay, Sophie; Warwick, Helen K.; Machesky, Laura M.

doi:10.1091/mbc.e02-06-0345

Cited by 55 publications

(56 citation statements)

References 57 publications

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“…This is supported by our finding of a high degree of colocalization between the active N-WASP species and the transferrin receptor-positive compartment, as well as the recent demonstration that in yeast the WASP homolog Las17 powers the actin polymerization-dependent motility of endosomes via activation of the Arp2/3 complex (9). The Arp2/3 complex is also essential for Golgi polarization in wound edge NIH 3T3 cells (26). In N-WASP-deficient cells, the transit of hemagglutinin (as a protein marker) from the endoplasmic reticulum to the Golgi apparatus is reduced by 40%, demonstrating a specific role for N-WASP as a regulator of vesicle traffic between the endoplasmic reticulum and the Golgi complex (48).…”

Section: Discussionmentioning

confidence: 99%

Spatially Distinct Binding of Cdc42 to PAK1 and N-WASP in Breast Carcinoma Cells

Parsons

Monypenny²,

Ameer-Beg

et al. 2005

Molecular and Cellular Biology

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While a significant amount is known about the biochemical signaling pathways of the Rho family GTPase Cdc42, a better understanding of how these signaling networks are coordinated in cells is required. In particular, the predominant subcellular sites where GTP-bound Cdc42 binds to its effectors, such as p21-activated kinase 1 (PAK1) and N-WASP, a homolog of the Wiskott-Aldritch syndrome protein, are still undetermined. Recent fluorescence resonance energy transfer (FRET) imaging experiments using activity biosensors show inconsistencies between the site of local activity of PAK1 or N-WASP and the formation of specific membrane protrusion structures in the cell periphery. The data presented here demonstrate the localization of interactions by using multiphoton time-domain fluorescence lifetime imaging microscopy (FLIM). Our data here establish that activated Cdc42 interacts with PAK1 in a nucleotide-dependent manner in the cell periphery, leading to Thr-423 phosphorylation of PAK1, particularly along the lengths of cell protrusion structures. In contrast, the majority of GFP-N-WASP undergoing FRET with Cy3-Cdc42 is localized within a transferrin receptor-and Rab11-positive endosomal compartment in breast carcinoma cells. These data reveal for the first time distinct spatial association patterns between Cdc42 and its key effector proteins controlling cytoskeletal remodeling.Cytoskeletal remodelling is a highly dynamic, tightly regulated process which drives the formation of cell protrusions necessary for cell movement. Members of the Rho family of GTPases regulate cytoskeletal dynamics by cycling between inactive GDP-bound and active GTP-bound states. Cdc42 activation results in a localized increase in actin polymerization and the formation of filopodia. Immediately downstream of Cdc42 are p21-activated kinase 1 (PAK1) as well as the Wiskott-Aldrich syndrome protein (WASP) family of proteins, both of which are activated through the binding of GTP Cdc42 via the Cdc42-and Rac-binding domain (CRIB or PBD). PAK1 has been reported to localize to regions of cytoskeletal assembly, and the activated form can induce cytoskeletal remodeling at the leading edge via downstream events such as microtubule growth and stathmin phosphorylation (11, 60). A close correlation has been found between the PAK1 activity state and the baseline invasiveness of human breast cancer cells as well as breast tumor grades (52). Paradoxically, the same work demonstrated that breast carcinoma cells expressing the GTPase binding-deficient H83L, H86L PAK1 mutant exhibited extensive membrane ruffling in the cell periphery. The lamellipodia formation and membrane ruffling observed in the presence of PAK1 were subsequently demonstrated by others to be independent of PAK catalytic activity (13). The relationship between the local level of PAK1 activity and the formation of specific subcellular structures is therefore unclear.The WASP-family proteins and the Arp2/3 complex are important nucleators of new actin filaments in response to signals causing cel...

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Section: Discussionmentioning

confidence: 99%

Spatially Distinct Binding of Cdc42 to PAK1 and N-WASP in Breast Carcinoma Cells

Parsons

Monypenny²,

Ameer-Beg

et al. 2005

Molecular and Cellular Biology

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“…In migrating astrocytes, the Cdc42-Par6-aPKC pathway controls the orientation of the Golgi, probably indirectly by modifying microtubule organization. By contrast, in NIH3T3 cells, regulation of the actin cytoskeleton by Scar2 and the Arp2/3 complex has been implicated in Golgi reorientation (Magdalena et al, 2003). Moreover, active Cdc42 binds directly to the coatomer complex and regulates vesicular trafficking between the endoplasmic reticulum and Golgi (Wu et al, 2000).…”

Section: Cdc42 Regulates Membrane Trafficmentioning

confidence: 98%

Cdc42 - the centre of polarity

Etienne-Manneville

2004

Journal of Cell Science

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683

642

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All cell types polarize, at least transiently, during division or to generate specialized shapes and functions. This capacity extends from yeast to mammals, and it is now clear that many features of the molecular mechanisms controlling polarization are conserved in all eukaryotic cells. At the centre of the action is Cdc42, a small GTPase of the Rho family. Its activity is precisely controlled both temporally and spatially, and this can be achieved by a wide variety of extracellular cues in multicellular organisms. Moreover, although the functional characteristics of cell polarity are extremely variable (depending on the cell type and the biological context), Cdc42 has an amazing capacity to co-ordinate the control of multiple signal transduction pathways.

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“…S4) in scratch wound assays. Scratch assays are commonly used to study the ability of cells to polarize and migrate into the wound with time (35,36). Cells expressing the C17orf37…”

Section: C17orf37 Prenylation Regulates Functional Response Of the Prmentioning

confidence: 99%

Prenylated C17orf37 Induces Filopodia Formation to Promote Cell Migration and Metastasis

Dasgupta

Cushman

Kpetemey

et al. 2011

Journal of Biological Chemistry

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Post-translational modification by covalent attachment of isoprenoid lipids (prenylation) regulates the functions and biological activities of several proteins implicated in the oncogenic transformation and metastatic progression of cancer. The largest group of prenylated proteins contains a CAAX motif at the C-terminal that serves as a substrate for a series of post-translational modifications that convert these otherwise hydrophilic proteins to lipidated proteins, thus facilitating membrane association. C17orf37 (chromosome 17 open reading frame 37), also known as C35/Rdx12/MGC14832, located in the 17q12 amplicon, is overexpressed in human cancer, and its expression correlates with the migratory and invasive phenotype of cancer cells. Here we show that C17orf37 contains a functional CAAX motif and is post-translationally modified by protein geranylgeranyltransferase-I (GGTase-I). Geranylgeranylation of C17orf37 at the CAAX motif facilitates association of the protein to the inner leaflet of plasma membrane, enhances migratory phenotype of cells by inducing increased filopodia formation, and potentiates directional migration. A prenylation-deficient mutant of C17orf37 is functionally inactive and fails to trigger dissemination of tail vein-injected cells in a mouse model of metastasis. These findings demonstrate that prenylation is required for the function of the C17orf37 protein in cancer cells and imply that the post-translational modification may functionally regulate metastatic progression of disease.

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Involvement of the Arp2/3 Complex and Scar2 in Golgi Polarity in Scratch Wound Models

Cited by 55 publications

References 57 publications

Spatially Distinct Binding of Cdc42 to PAK1 and N-WASP in Breast Carcinoma Cells

Spatially Distinct Binding of Cdc42 to PAK1 and N-WASP in Breast Carcinoma Cells

Cdc42 - the centre of polarity

Prenylated C17orf37 Induces Filopodia Formation to Promote Cell Migration and Metastasis

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