scholarly journals Endosomal WASH and exocyst complexes control exocytosis of MT1-MMP at invadopodia

2013 ◽  
Vol 203 (6) ◽  
pp. 1063-1079 ◽  
Author(s):  
Pedro Monteiro ◽  
Carine Rossé ◽  
Antonio Castro-Castro ◽  
Marie Irondelle ◽  
Emilie Lagoutte ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Carcinoma Cells ◽  
General Mechanism ◽  
Pericellular Matrix ◽  
Matrix Degradation ◽  
Late Endosomes ◽  
The Matrix ◽  
Membrane Type ◽  
Syndrome Protein

Remodeling of the extracellular matrix by carcinoma cells during metastatic dissemination requires formation of actin-based protrusions of the plasma membrane called invadopodia, where the trans-membrane type 1 matrix metalloproteinase (MT1-MMP) accumulates. Here, we describe an interaction between the exocyst complex and the endosomal Arp2/3 activator Wiskott-Aldrich syndrome protein and Scar homolog (WASH) on MT1-MMP–containing late endosomes in invasive breast carcinoma cells. We found that WASH and exocyst are required for matrix degradation by an exocytic mechanism that involves tubular connections between MT1-MMP–positive late endosomes and the plasma membrane in contact with the matrix. This ensures focal delivery of MT1-MMP and supports pericellular matrix degradation and tumor cell invasion into different pathologically relevant matrix environments. Our data suggest a general mechanism used by tumor cells to breach the basement membrane and for invasive migration through fibrous collagen-enriched tissues surrounding the tumor.

scholarly journals N-WASP coordinates the delivery and F-actin–mediated capture of MT1-MMP at invasive pseudopods

2012 ◽  
Vol 199 (3) ◽  
pp. 527-544 ◽  
Author(s):  
Xinzi Yu ◽  
Tobias Zech ◽  
Laura McDonald ◽  
Esther Garcia Gonzalez ◽  
Ang Li ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Extracellular Matrix ◽  
Plasma Membrane ◽  
Tumor Cells ◽  
Matrix Metalloproteases ◽  
Matrix Remodeling ◽  
Dense Matrix ◽  
Matrix Degradation ◽  
Late Endosomes ◽  
Syndrome Protein

Metastasizing tumor cells use matrix metalloproteases, such as the transmembrane collagenase MT1-MMP, together with actin-based protrusions, to break through extracellular matrix barriers and migrate in dense matrix. Here we show that the actin nucleation–promoting protein N-WASP (Neural Wiskott-Aldrich syndrome protein) is up-regulated in breast cancer, and has a pivotal role in mediating the assembly of elongated pseudopodia that are instrumental in matrix degradation. Although a role for N-WASP in invadopodia was known, we now show how N-WASP regulates invasive protrusion in 3D matrices. In actively invading cells, N-WASP promoted trafficking of MT1-MMP into invasive pseudopodia, primarily from late endosomes, from which it was delivered to the plasma membrane. Upon MT1-MMP’s arrival at the plasma membrane in pseudopodia, N-WASP stabilized MT1-MMP via direct tethering of its cytoplasmic tail to F-actin. Thus, N-WASP is crucial for extension of invasive pseudopods into which MT1-MMP traffics and for providing the correct cytoskeletal framework to couple matrix remodeling with protrusive invasion.

scholarly journals The interaction of IQGAP1 with the exocyst complex is required for tumor cell invasion downstream of Cdc42 and RhoA

2008 ◽  
Vol 181 (6) ◽  
pp. 985-998 ◽  
Author(s):  
Mika Sakurai-Yageta ◽  
Chiara Recchi ◽  
Gaëlle Le Dez ◽  
Jean-Baptiste Sibarita ◽  
Laurent Daviet ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Surface Membrane ◽  
Carcinoma Cells ◽  
Matrix Degradation ◽  
Vesicle Tethering ◽  
Exocyst Complex ◽  
Membrane Protrusions ◽  
Actin Regulatory Proteins ◽  
Membrane Type

Invadopodia are actin-based membrane protrusions formed at contact sites between invasive tumor cells and the extracellular matrix with matrix proteolytic activity. Actin regulatory proteins participate in invadopodia formation, whereas matrix degradation requires metalloproteinases (MMPs) targeted to invadopodia. In this study, we show that the vesicle-tethering exocyst complex is required for matrix proteolysis and invasion of breast carcinoma cells. We demonstrate that the exocyst subunits Sec3 and Sec8 interact with the polarity protein IQGAP1 and that this interaction is triggered by active Cdc42 and RhoA, which are essential for matrix degradation. Interaction between IQGAP1 and the exocyst is necessary for invadopodia activity because enhancement of matrix degradation induced by the expression of IQGAP1 is lost upon deletion of the exocyst-binding site. We further show that the exocyst and IQGAP1 are required for the accumulation of cell surface membrane type 1 MMP at invadopodia. Based on these results, we propose that invadopodia function in tumor cells relies on the coordination of cytoskeletal assembly and exocytosis downstream of Rho guanosine triphosphatases.

scholarly journals Microenvironmental Regulation of Membrane Type 1 Matrix Metalloproteinase Activity in Ovarian Carcinoma Cells via Collagen-induced EGR1 Expression

2006 ◽  
Vol 282 (7) ◽  
pp. 4924-4931 ◽  
Author(s):  
Maria V. Barbolina ◽  
Brian P. Adley ◽  
Edgardo V. Ariztia ◽  
Yueying Liu ◽  
M. Sharon Stack
Keyword(s):  
Matrix Metalloproteinase ◽  
Ovarian Carcinoma ◽  
Three Dimensional ◽  
Carcinoma Cells ◽  
Collagenolytic Activity ◽  
Type I ◽  
Collagen Gels ◽  
Ovarian Carcinoma Cells ◽  
Membrane Type

Late stage ovarian cancer is characterized by disseminated intraperitoneal metastasis as secondary lesions anchor in the type I and III collagen-rich submesothelial matrix. Ovarian carcinoma cells preferentially adhere to interstitial collagen, and collagen-induced integrin clustering up-regulates the expression of the transmembrane collagenase membrane type 1 matrix metalloproteinase (MT1-MMP). Collagenolytic activity is important in intraperitoneal metastasis, potentiating invasion through the mesothelial cell layer and colonization of the submesothelial collagen-rich matrix. The objective of this study was to elucidate a potential mechanistic link between collagen adhesion and MT1-MMP expression. Our results indicate that culturing cells on three-dimensional collagen gels, but not thin layer collagen or synthetic threedimensional hydrogels, results in rapid induction of the transcription factor EGR1. Integrin signaling through a SRC kinase-dependent pathway is necessary for EGR1 induction. Silencing of EGR1 expression using small interfering RNA abrogated collagen-induced MT1-MMP expression and inhibited cellular invasion of three-dimensional collagen gels. These data support a model for intraperitoneal metastasis wherein collagen adhesion and clustering of collagen binding integrins activates integrin-mediated signaling via SRC kinases to induce expression of EGR1, resulting in transcriptional activation of the MT1-MMP promoter and subsequent MT1-MMP-catalyzed collagen invasion. This model highlights the role of unique interactions between ovarian carcinoma cells and interstitial collagens in the ovarian tumor microenvironment in inducing gene expression changes that potentiate intraperitoneal metastatic progression.

scholarly journals A Ca2+-stimulated exosome release pathway in cancer cells is regulated by Munc13-4

2018 ◽  
Vol 217 (8) ◽  
pp. 2877-2890 ◽  
Author(s):  
Scott W. Messenger ◽  
Sang Su Woo ◽  
Zhongze Sun ◽  
Thomas F.J. Martin
Keyword(s):  
Extracellular Matrix ◽  
Cancer Cells ◽  
Multivesicular Bodies ◽  
Matrix Degradation ◽  
Cellular Mechanisms ◽  
Trafficking Pathway ◽  
Acute Elevation ◽  
Membrane Type ◽  
Dependent Pathway

Cancer cells secrete copious amounts of exosomes, and elevated intracellular Ca2+ is critical for tumor progression and metastasis, but the underlying cellular mechanisms are unknown. Munc13-4 is a Ca2+-dependent SNAP receptor– and Rab-binding protein required for Ca2+-dependent membrane fusion. Here we show that acute elevation of Ca2+ in cancer cells stimulated a fivefold increase in CD63+, CD9+, and ALIX+ exosome release that was eliminated by Munc13-4 knockdown and not restored by Ca2+ binding–deficient Munc13-4 mutants. Direct imaging of CD63-pHluorin exosome release confirmed its Munc13-4 dependence. Depletion of Munc13-4 in highly aggressive breast carcinoma MDA-MB-231 cells reduced the size of CD63+ multivesicular bodies (MVBs), indicating a role for Munc13-4 in MVB maturation. Munc13-4 used a Rab11-dependent trafficking pathway to generate MVBs competent for exosome release. Membrane type 1 matrix metalloproteinase trafficking to MVBs by a Rab11-dependent pathway was also Munc13-4 dependent, and Munc13-4 depletion reduced extracellular matrix degradation. These studies identify a novel Ca2+- and Munc13-4-dependent pathway that underlies increased exosome release by cancer cells.

scholarly journals MT1-MMP recruits the ER-Golgi SNARE Bet1 for efficient MT1-MMP transport to the plasma membrane

2019 ◽  
Vol 218 (10) ◽  
pp. 3355-3371 ◽  
Author(s):  
Takuya Miyagawa ◽  
Kana Hasegawa ◽  
Yoko Aoki ◽  
Takuya Watanabe ◽  
Yuka Otagiri ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Plasma Membrane ◽  
Intracellular Transport ◽  
Early Stage ◽  
Snare Complex ◽  
Invasion And Metastasis ◽  
Golgi Transport ◽  
Syntaxin 4 ◽  
Membrane Type

Metastasis is a major cause of cancer-related death. Membrane type 1–matrix metalloproteinase (MT1-MMP) is a critical protease for local invasion and metastasis. MT1-MMP is synthesized in the endoplasmic reticulum (ER) and transported in vesicles to invadopodia, specialized subdomains of the plasma membrane, through secretory and endocytic recycling pathways. The molecular mechanism underlying intracellular transport of MT1-MMP has been extensively studied, but is not fully understood. We show that MT1-MMP diverts the SNARE Bet1 from its function in ER-Golgi transport, to promote MT1-MMP trafficking to the cell surface, likely to invadopodia. In invasive cells, Bet1 is localized in MT1-MMP–positive endosomes in addition to the Golgi apparatus, and forms a novel SNARE complex with syntaxin 4 and endosomal SNAREs. MT1-MMP may also use Bet1 for its export from raft-like structures in the ER. Our results suggest the recruitment of Bet1 at an early stage after MT1-MMP expression promotes the exit of MT1-MMP from the ER and its efficient transport to invadopodia.

scholarly journals ARF6–JIP3/4 regulate endosomal tubules for MT1-MMP exocytosis in cancer invasion

2015 ◽  
Vol 211 (2) ◽  
pp. 339-358 ◽  
Author(s):  
Valentina Marchesin ◽  
Antonio Castro-Castro ◽  
Catalina Lodillinsky ◽  
Alessia Castagnino ◽  
Joanna Cyrta ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Tumor Cells ◽  
Cell Biology ◽  
Breast Cancer Cells ◽  
Triple Negative ◽  
Breast Cancers ◽  
Interacting Protein ◽  
Membrane Type ◽  
Syndrome Protein

Invasion of cancer cells into collagen-rich extracellular matrix requires membrane-tethered membrane type 1–matrix metalloproteinase (MT1-MMP) as the key protease for collagen breakdown. Understanding how MT1-MMP is delivered to the surface of tumor cells is essential for cancer cell biology. In this study, we identify ARF6 together with c-Jun NH2-terminal kinase–interacting protein 3 and 4 (JIP3 and JIP4) effectors as critical regulators of this process. Silencing ARF6 or JIP3/JIP4 in breast tumor cells results in MT1-MMP endosome mispositioning and reduces MT1-MMP exocytosis and tumor cell invasion. JIPs are recruited by Wiskott-Aldrich syndrome protein and scar homologue (WASH) on MT1-MMP endosomes on which they recruit dynein–dynactin and kinesin-1. The interaction of plasma membrane ARF6 with endosomal JIPs coordinates dynactin–dynein and kinesin-1 activity in a tug-of-war mechanism, leading to MT1-MMP endosome tubulation and exocytosis. In addition, we find that ARF6, MT1-MMP, and kinesin-1 are up-regulated in high-grade triple-negative breast cancers. These data identify a critical ARF6–JIP–MT1-MMP–dynein–dynactin–kinesin-1 axis promoting an invasive phenotype of breast cancer cells.

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