scholarly journals Nance-Horan Syndrome-like 1 protein negatively regulates Scar/WAVE-Arp2/3 activity and inhibits lamellipodia stability and cell migration

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Ah-Lai Law ◽  
Shamsinar Jalal ◽  
Tommy Pallett ◽  
Fuad Mosis ◽  
Ahmad Guni ◽  
...  
Keyword(s):  
Cell Migration ◽  
Binding Sites ◽  
Leading Edge ◽  
Mesenchymal Cell ◽  
Negative Regulators ◽  
Binding Partner ◽  
Direct Binding ◽  
The Stability ◽  
Wave Complex ◽  
Lamellipodia Formation

AbstractCell migration is important for development and its aberrant regulation contributes to many diseases. The Scar/WAVE complex is essential for Arp2/3 mediated lamellipodia formation during mesenchymal cell migration and several coinciding signals activate it. However, so far, no direct negative regulators are known. Here we identify Nance-Horan Syndrome-like 1 protein (NHSL1) as a direct binding partner of the Scar/WAVE complex, which co-localise at protruding lamellipodia. This interaction is mediated by the Abi SH3 domain and two binding sites in NHSL1. Furthermore, active Rac binds to NHSL1 at two regions that mediate leading edge targeting of NHSL1. Surprisingly, NHSL1 inhibits cell migration through its interaction with the Scar/WAVE complex. Mechanistically, NHSL1 may reduce cell migration efficiency by impeding Arp2/3 activity, as measured in cells using a Arp2/3 FRET-FLIM biosensor, resulting in reduced F-actin density of lamellipodia, and consequently impairing the stability of lamellipodia protrusions.

scholarly journals Nance-Horan Syndrome-like 1 protein negatively regulates Scar/WAVE-Arp2/3 activity and inhibits lamellipodia stability and cell migration

2020 ◽  
Author(s):  
Ah-Lai Law ◽  
Shamsinar Jalal ◽  
Fuad Mosis ◽  
Tommy Pallett ◽  
Ahmad Guni ◽  
...  
Keyword(s):  
Cell Migration ◽  
Binding Sites ◽  
Leading Edge ◽  
Mesenchymal Cell ◽  
Negative Regulators ◽  
Binding Partner ◽  
Direct Binding ◽  
The Stability ◽  
Wave Complex ◽  
Lamellipodia Formation

AbstractCell migration is important for development and its aberrant regulation contributes to many diseases. The Scar/WAVE complex is essential for Arp2/3 mediated lamellipodia formation during mesenchymal cell migration and several coinciding signals activate it. However, so far, no direct negative regulators are known. We have identified Nance-Horan Syndrome-like 1 protein (NHSL1) as a novel, direct binding partner of the Scar/WAVE complex, which co-localise at protruding lamellipodia. This interaction is mediated by the Abi SH3 domain and two binding sites in NHSL1. Furthermore, active Rac binds to NHSL1 at two regions that mediate leading edge targeting of NHSL1 suggesting that Rac recruits NHSL1. Surprisingly, NHSL1 inhibits cell migration through its interaction with the Scar/WAVE complex. Mechanistically, NHSL1 may reduce cell migration efficiency by impeding Arp2/3 activity, as measured in cells using a novel Arp2/3 FRET-FLIM biosensor, resulting in reduced F-actin content of lamellipodia, and consequently impairing the stability of lamellipodia protrusions.

Control of stable lamellipodia formation by expression of cell surface beta 1,4-galactosyltransferase cytoplasmic domains

1994 ◽  
Vol 107 (9) ◽  
pp. 2535-2545 ◽  
Author(s):  
P.A. Appeddu ◽  
B.D. Shur
Keyword(s):  
Growth Factor ◽  
Cell Surface ◽  
Binding Sites ◽  
Mesenchymal Cell ◽  
Fiber Formation ◽  
Platelet Derived Growth Factor ◽  
Specific Stimulus ◽  
3T3 Fibroblasts ◽  
A Cell ◽  
Lamellipodia Formation

Mesenchymal cell migration on basal lamina is mediated, in part, by the binding of cell surface beta 1,4-galactosyltransferase (GalTase) to specific N-linked oligosaccharides in the E8 domain of laminin. On migrating cells, surface GalTase is anchored to the cytoskeleton; when GalTase is prevented from associating with the cytoskeleton, lamellipodia formation and subsequent migration are inhibited. To define better the involvement of GalTase-cytoskeleton interactions in cell motility, we examined the lamellipodia formation, polarity and migratory behavior of stably transfected 3T3 fibroblasts expressing increased or decreased levels of GalTase capable of interacting with the cytoskeleton. Initially, the motile behavior of individual cells was quantified in the absence of exogenous stimuli. Cells that overexpress GalTase binding sites for the cytoskeleton changed their polarity more frequently and translocated more erratically than did control cells when assayed on laminin substrata. These differences were not observed, however, when cells were plated on fibronectin, which does not contain binding sites for surface GalTase. GalTase-transfected cells were also assayed for their ability to polarize in response to a specific stimulus. In this case, the ability of a cell to reorient towards a gradient of platelet-derived growth factor was found to be directly proportional to the amount of GalTase associated with the cytoskeleton. Differences in response to platelet-derived growth factor were not due to differences in growth factor binding. Indirect immunofluorescence showed that altering the level of GalTase did not affect the ventrally distributed pool of GalTase stably associated with the cytoskeleton; however, stress fiber formation was inhibited. Thus, increasing surface GalTase binding sites for the cytoskeleton leads to erratic, multipolar behavior in the absence of any vectorial stimulus, but the ability to form a functional lamellipodium in response to a stimulus is dependent upon the amount of surface GalTase associated with the cytoskeleton. Apparently, cells are able to regulate cytoskeletal assembly and lamellipodial stability by altering the expression and/or affinity of appropriate matrix receptors, such as GalTase, and their corresponding binding sites in the cytoskeleton.

scholarly journals Rac1 and Stathmin but Not EB1 Are Required for Invasion of Breast Cancer Cells in Response to IGF-I

2011 ◽  
Vol 2011 ◽  
pp. 1-9 ◽  
Author(s):  
Shigeru Morimura ◽  
Kazuhide Takahashi
Keyword(s):  
Breast Cancer ◽  
Cell Migration ◽  
Cell Invasion ◽  
Breast Cancer Cells ◽  
Regulatory Protein ◽  
Leading Edge ◽  
Mcf7 Cells ◽  
Igf I ◽  
Interfering Rna ◽  
Lamellipodia Formation

Cell migration is considered necessary for the invasion that accompanies the directional formation of the cellular protrusions termed lamellipodia. In invasive breast cancer MDA-MB-231 cells, lamellipodia formation is preceded by translocation of the actin cytoskeletal regulatory protein WAVE2 to the leading edge. WAVE2 translocation and lamellipodia formation require many signaling molecules, including PI3K, Rac1, Pak1, IRSp53, stathmin, and EB1, but whether these molecules are necessary for invasion remains unclear. In noninvasive breast cancer MCF7 cells, no lamellipodia were induced by IGF-I, whereas in MDA-MB-231 cells, Rac1, stathmin, and EB1 were overexpressed. Depletion of Rac1 or stathmin by small interfering RNA abrogated the IGF-I-induced invasion of MDA-MB-231 cells; however, depletion of EB1 did not, indicating the necessity of Rac1 and stathmin but not EB1 for invasion. The signaling pathway leading to cell invasion may not be identical but shares some common molecules, leading to cell migration through lamellipodia formation.

scholarly journals WAVE3-mediated Cell Migration and Lamellipodia Formation Are Regulated Downstream of Phosphatidylinositol 3-Kinase

2005 ◽  
Vol 280 (23) ◽  
pp. 21748-21755 ◽  
Author(s):  
Khalid Sossey-Alaoui ◽  
Xiurong Li ◽  
Tamara A. Ranalli ◽  
John K. Cowell
Keyword(s):  
Cell Migration ◽  
Three Dimensional ◽  
Cell Movement ◽  
Leading Edge ◽  
Sh2 Domain ◽  
Regulatory Subunit ◽  
Phosphatidylinositol 3 Kinase ◽  
Cytoskeleton Reorganization ◽  
Lamellipodia Formation ◽  
Phosphatidylinositol 3

WAVE3 is a member of the WASP/WAVE family of protein effectors of actin reorganization and cell movement. The precise role of WAVE3 in cell migration and its regulation, however, have not been elucidated. Here we show that endogenous WAVE3 was found to be concentrated in the lamellipodia at the leading edge of migrating MDA-MB-231 cells. Platelet-derived growth factor (PDGF) treatment induced lamellipodia formation as well as two-dimensional migration of cells in the wound-closure assay and chemotactic migration toward PDGF in three-dimensional migration chambers. Knockdown of WAVE3 expression by RNA interference prevented the PDGF-induced lamellipodia formation and cell migration. Treatment of cells with LY294002, an inhibitor of phosphatidylinositol 3-kinase (PI3K), also abrogated the PDGF-induced lamellipodia formation and cell migration, suggesting that PI3K may be required for WAVE3 activity. WAVE3 and the PI3K regulatory subunit, p85, were found to interact in a yeast two-hybrid screen, which was confirmed through co-immunoprecipitation. The WAVE3-p85 interaction was mediated by the N-terminal region of WAVE3 and the C-terminal SH2 domain of p85. These results imply that the WAVE3-mediated migration in MDA-MB-231 cells via lamellipodia formation is activated downstream of PI3K and induced by PDGF. The findings of the WAVE3-p85 partnership also suggest a potential regulatory role for p85 in WAVE3-dependent actin-cytoskeleton reorganization and cell migration.

scholarly journals CYFIP2 containing WAVE complexes inhibit cell migration

2020 ◽  
Author(s):  
Anna Polesskaya ◽  
Arthur Boutillon ◽  
Yanan Wang ◽  
Marc Lavielle ◽  
Sophie Vacher ◽  
...  
Keyword(s):  
Cell Migration ◽  
Mammary Epithelial Cells ◽  
Leading Edge ◽  
Good Prognosis ◽  
Mammary Epithelial ◽  
Breast Cancer Patients ◽  
Loss Of Function ◽  
A Cell ◽  
Wave Complex

ABSTRACTBranched actin networks polymerized by the Arp2/3 complex are critical for cell migration. The WAVE complex is the major Arp2/3 activator at the leading edge of migrating cells. However, multiple distinct WAVE complexes can be assembled in a cell, due to the combinatorial complexity of paralogous subunits. When systematically analyzing the contribution of each WAVE complex subunit to the metastasis-free survival of breast cancer patients, we found that overexpression of the CYFIP2 subunit was surprisingly associated with good prognosis. Gain and loss of function experiments in transformed and untransformed mammary epithelial cells revealed that cell migration was always inversely related to CYFIP2 levels. The role of CYFIP2 was systematically opposite to the role of the paralogous subunit CYFIP1 or of the NCKAP1 subunit. The specific CYFIP2 function in inhibiting cell migration was related to its unique ability to down-regulate classical pro-migratory WAVE complexes. The anti-migratory function of CYFIP2 was also revealed in migration of prechordal plate cells during gastrulation of the zebrafish embryo, indicating that the unique function of CYFIP2 is critically important in both physiological and pathophysiological migrations.

scholarly journals ERBB3-dependent AKT and ERK pathways are essential for atrioventricular cushion development in mouse embryos

PLoS ONE ◽  
2021 ◽  
Vol 16 (10) ◽  
pp. e0259426
Author(s):  
Kyoungmi Kim ◽  
Daekee Lee
Keyword(s):  
Binding Sites ◽  
Heart Development ◽  
Mesenchymal Cell ◽  
Embryonic Heart ◽  
Regulatory Subunit ◽  
Akt Pathway ◽  
Erk Activation ◽  
Binding Partner ◽  
Downstream Signaling ◽  
Heart Physiology

ERBB family members and their ligands play an essential role in embryonic heart development and adult heart physiology. Among them, ERBB3 is a binding partner of ERBB2; the ERBB2/3 complex mediates downstream signaling for cell proliferation. ERBB3 has seven consensus binding sites to the p85 regulatory subunit of PI3K, which activates the downstream AKT pathway, leading to the proliferation of various cells. This study generated a human ERBB3 knock-in mouse expressing a mutant ERBB3 whose seven YXXM p85 binding sites were replaced with YXXA. Erbb3 knock-in embryos exhibited lethality between E12.5 to E13.5, and showed a decrease in mesenchymal cell numbers and density in AV cushions. We determined that the proliferation of mesenchymal cells in the atrioventricular (AV) cushion in Erbb3 knock-in mutant embryos was temporarily reduced due to the decrease of AKT and ERK1/2 phosphorylation. Overall, our results suggest that AKT/ERK activation by the ERBB3-dependent PI3K signaling is crucial for AV cushion morphogenesis during embryonic heart development.

scholarly journals Activation of Gαi3 triggers cell migration via regulation of GIV

2008 ◽  
Vol 182 (2) ◽  
pp. 381-393 ◽  
Author(s):  
Pradipta Ghosh ◽  
Mikel Garcia-Marcos ◽  
Scott J. Bornheimer ◽  
Marilyn G. Farquhar
Keyword(s):  
Signal Transduction ◽  
Wound Healing ◽  
Cell Migration ◽  
Mammalian Cells ◽  
Leading Edge ◽  
Akt Signaling ◽  
Actin Remodeling ◽  
Tumor Cell Migration ◽  
Binding Partner ◽  
Macrophage Chemotaxis

During migration, cells must couple direction sensing to signal transduction and actin remodeling. We previously identified GIV/Girdin as a Gαi3 binding partner. We demonstrate that in mammalian cells Gαi3 controls the functions of GIV during cell migration. We find that Gαi3 preferentially localizes to the leading edge and that cells lacking Gαi3 fail to polarize or migrate. A conformational change induced by association of GIV with Gαi3 promotes Akt-mediated phosphorylation of GIV, resulting in its redistribution to the plasma membrane. Activation of Gαi3 serves as a molecular switch that triggers dissociation of Gβγ and GIV from the Gi3–GIV complex, thereby promoting cell migration by enhancing Akt signaling and actin remodeling. Gαi3–GIV coupling is essential for cell migration during wound healing, macrophage chemotaxis, and tumor cell migration, indicating that the Gαi3–GIV switch serves to link direction sensing from different families of chemotactic receptors to formation of the leading edge during cell migration.

scholarly journals The SCAR/WAVE complex is necessary for proper regulation of traction stresses during amoeboid motility

2011 ◽  
Vol 22 (21) ◽  
pp. 3995-4003 ◽  
Author(s):  
Effie Bastounis ◽  
Ruedi Meili ◽  
Baldomero Alonso-Latorre ◽  
Juan C. del Álamo ◽  
Juan C. Lasheras ◽  
...  
Keyword(s):  
Cell Migration ◽  
Actin Polymerization ◽  
Leading Edge ◽  
Wild Type ◽  
Amoeboid Motility ◽  
Elastic Substrates ◽  
Random Manner ◽  
Motility Cycle ◽  
Wave Complex

Cell migration requires a tightly regulated, spatiotemporal coordination of underlying biochemical pathways. Crucial to cell migration is SCAR/WAVE–mediated dendritic F-actin polymerization at the cell's leading edge. Our goal is to understand the role the SCAR/WAVE complex plays in the mechanics of amoeboid migration. To this aim, we measured and compared the traction stresses exerted by Dictyostelium cells lacking the SCAR/WAVE complex proteins PIR121 (pirA−) and SCAR (scrA−) with those of wild-type cells while they were migrating on flat, elastic substrates. We found that, compared to wild type, both mutant strains exert traction stresses of different strengths that correlate with their F-actin levels. In agreement with previous studies, we found that wild-type cells migrate by repeating a motility cycle in which the cell length and strain energy exerted by the cells on their substrate vary periodically. Our analysis also revealed that scrA− cells display an altered motility cycle with a longer period and a lower migration velocity, whereas pirA− cells migrate in a random manner without implementing a periodic cycle. We present detailed characterization of the traction-stress phenotypes of the various cell lines, providing new insights into the role of F-actin polymerization in regulating cell–substratum interactions and stresses required for motility.

scholarly journals Rab5 Induces Rac-independent Lamellipodia Formation and Cell Migration

1999 ◽  
Vol 10 (10) ◽  
pp. 3239-3250 ◽  
Author(s):  
Marcel Spaargaren ◽  
Johannes L. Bos
Keyword(s):  
Cell Migration ◽  
Actin Cytoskeleton ◽  
Actin Polymerization ◽  
Strong Support ◽  
Leading Edge ◽  
Vesicle Fusion ◽  
Dominant Negative ◽  
Cytoskeleton Reorganization ◽  
Endocytic Vesicles ◽  
Lamellipodia Formation

Rab5 is a regulatory GTPase of vesicle docking and fusion that is involved in receptor-mediated endocytosis and pinocytosis. Introduction of active Rab5 in cells stimulates the rate of endocytosis and vesicle fusion, resulting in the formation of large endocytic vesicles, whereas dominant negative Rab5 inhibits vesicle fusion. Here we show that introduction of active Rab5 in fibroblasts also induced reorganization of the actin cytoskeleton but not of microtubule filaments, resulting in prominent lamellipodia formation. The Rab5-induced lamellipodia formation did not require activation of PI3-K or the GTPases Ras, Rac, Cdc42, or Rho, which are all strongly implicated in cytoskeletal reorganization. Furthermore, lamellipodia formation by insulin, Ras, or Rac was not affected by expression of dominant negative Rab5. In addition, cells expressing active Rab5 displayed a dramatic stimulation of cell migration, with the lamellipodia serving as the leading edge. Both lamellipodia formation and cell migration were dependent on actin polymerization but not on microtubules. These results demonstrate that Rab5 induces lamellipodia formation and cell migration and that the Rab5-induced lamellipodia formation occurs by a novel mechanism independent of, and distinct from, PI3-K, Ras, or Rho-family GTPases. Thus, Rab5 can control not only endocytosis but also actin cytoskeleton reorganization and cell migration, which provides strong support for an intricate relationship between these processes.

scholarly journals Cytoskeletal Mechanics Regulating Amoeboid Cell Locomotion

2014 ◽  
Vol 66 (5) ◽  
Author(s):  
Begoña Álvarez-González ◽  
Effie Bastounis ◽  
Ruedi Meili ◽  
Juan C. del Álamo ◽  
Richard Firtel ◽  
...  
Keyword(s):  
Cell Migration ◽  
Traction Force ◽  
Leading Edge ◽  
Cell Types ◽  
Traction Forces ◽  
Amoeboid Cell ◽  
Periodic Movement ◽  
Wide Range ◽  
Motility Cycle ◽  
Wave Complex

Migrating cells exert traction forces when moving. Amoeboid cell migration is a common type of cell migration that appears in many physiological and pathological processes and is performed by a wide variety of cell types. Understanding the coupling of the biochemistry and mechanics underlying the process of migration has the potential to guide the development of pharmacological treatment or genetic manipulations to treat a wide range of diseases. The measurement of the spatiotemporal evolution of the traction forces that produce the movement is an important aspect for the characterization of the locomotion mechanics. There are several methods to calculate the traction forces exerted by the cells. Currently the most commonly used ones are traction force microscopy methods based on the measurement of the deformation induced by the cells on elastic substrate on which they are moving. Amoeboid cells migrate by implementing a motility cycle based on the sequential repetition of four phases. In this paper, we review the role that specific cytoskeletal components play in the regulation of the cell migration mechanics. We investigate the role of specific cytoskeletal components regarding the ability of the cells to perform the motility cycle effectively and the generation of traction forces. The actin nucleation in the leading edge of the cell, carried by the ARP2/3 complex activated through the SCAR/WAVE complex, has shown to be fundamental to the execution of the cyclic movement and to the generation of the traction forces. The protein PIR121, a member of the SCAR/WAVE complex, is essential to the proper regulation of the periodic movement and the protein SCAR, also included in the SCAR/WAVE complex, is necessary for the generation of the traction forces during migration. The protein Myosin II, an important F-actin cross-linker and motor protein, is essential to cytoskeletal contractility and to the generation and proper organization of the traction forces during migration.

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