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Cited by 912Extracellular vesicles in diseaseCellular transport and secretionCaveolin-1 and cellular processes

ARF6-Regulated Shedding of Tumor Cell-Derived Plasma Membrane Microvesicles

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Muralidharan-Chari, Vandhana, Clancy, James, Plou, Carolyn, Romao, Maryse, Chavrier, Philippe, Raposo, Graca, D'Souza-Schorey, Crislyn·2009·Current Biology
DOI 10.1016/j.cub.2009.09.059PMC PMC3150487
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Abstract

Background Increased mitogen-activated protein kinase (MAPK) signaling, small GTPase activation, cytoskeletal rearrangements, and the directed targeting of proteases to sites of extracellular matrix degradation all accompany the process of tumor cell invasion. Several studies have implicated the small GTP-binding protein ARF6 in tumor cell invasion, although the molecular basis by which ARF6 facilitates this process is unclear. Results We show that the ARF6 GTP/GDP cycle regulates the release of protease-loaded plasma membrane-derived microvesicles from tumor cells into the surrounding environment. To enable microvesicle shedding, ARF6-GTP-dependent activation of phospholipase D promotes the recruitment of the extracellular signal-regulated kinase (ERK) to the plasma membrane where, in turn, ERK phosphorylates and activates myosin light-chain kinase (MLCK). MLCK-mediated MLC phosphorylation is required for microvesicle release. Inhibition of ARF6 activation is accompanied by PKC-mediated phosphorylation of MLC, which blocks microvesicle shedding. Protein cargo appears to be selectively sorted into microvesicles, and adhesion to the extracellular matrix (ECM) is facilitated by microvesicle-associated integrin receptors. Conclusions Microvesicle shedding in tumor cells occurs via an actomyosin-based membrane abscission mechanism that is regulated by nucleotide cycling on ARF6. Microvesicle shedding appears to release selected cellular components, particularly those involved in cell adhesion and motility, into the surrounding environment. These findings suggest that ARF6 activation and the proteolytic activities of microvesicles, both of which are thought to correlate directly with tumor progression, could potentially serve as biomarkers for disease.

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  • Actin-myosin interactions appear critical to formation of all four EV types, and phosphatidylserine translocation to the outer membrane is common to apoptotic bodies and microvesicles.

    “For instance, actin-myosin interactions appear critical to the formation of all four types of EVs [66, 85, 100, 115].”

    From Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies (2013) · cites this paper as [66]

    The Results section ('ERK facilitates microvesicle shedding by phosphorylating myosin light chain') and Discussion support an actomyosin-dependent mechanism for microvesicle release. They do not establish that actin–myosin interactions are critical to formation of all four EV types. The Results also show phosphatidylserine externalization on microvesicles, but do not establish the stated comparison with apoptotic bodies. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • ARF6 overexpression increases microvesicle secretion without significantly altering exosome-sized vesicle secretion, supporting that microvesicle and exosome biogenesis are distinct.

    “In a melanoma model, overexpression of a rho family member, GTP-binding protein ADP-ribosylation factor 6 (ARF6), results in increased microvesicle secretion.”

    From Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies (2013) · cites this paper as [66]

    The Results section (‘Phenotypic variations of LOXARF6-GDP and LOXARF6-GTP cell lines’; page unavailable) reports significantly increased vesicle shedding from cells expressing active ARF6 compared with parental cells. The Introduction identifies ARF6 as belonging to the ARF family, not the Rho family as the printed citing sentence states. The Results distinguish the isolated microvesicles from exosomes by size and sedimentation, but do not establish the quote’s additional claim that exosome secretion was not significantly altered. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • CD63 and CD9 are potential exosome markers; ARF6 and VAMP3 are proposed microvesicle markers; TSP and C3b are accepted apoptotic body markers; Gag protein may be an RLP marker.

    “CD63 and CD9 are potential markers of exosomes [22, 117]. Markers of microvesicles are less well established though ARF6 and VCAMP3 are recently proposed [66]. TSP and C3b are generally accepted markers of apoptotic bodies. The RLPs are less well studied though Gag protein may be a marker for this group of EVs (Table 1).”

    From Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies (2013) · cites this paper as [66]

  • Microvesicle formation involves phosphatidylserine translocation to the outer membrane leaflet and completion by actin-myosin cytoskeletal contraction.

    “Membrane budding/vesicle formation is induced by translocation of phosphatidylserine to the outer-membrane leaflet [62, 65]. The budding process is completed through contraction of cytoskeletal structures by actin-myosin interactions [66, 67].”

    From Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies (2013) · cites this paper as [66]

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Cited by

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Authors
Muralidharan-Chari, V., Clancy, J., Plou, C., Romao, M., Chavrier, P., Raposo, G., & D'Souza-Schorey, C.
Affiliations
  • University of Notre Dame (US)
  • Institut Curie (FR)
ORCID
Venue
Current Biology
Publisher
Elsevier BV
Published
2009-12
Volume
19
Issue
22
Pages
1875-1885
ISSN
0960-9822 (ISSN-L)
DOI
10.1016/j.cub.2009.09.059
PMC
PMC3150487
OpenAlex
W2009649000
Cited by
912
References
1
Citations by year
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FWCI
5.91
Type
journal article
Language
en
Open access
Licence
Locations
  • Current Biology (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Science Foundation
  • National Cancer Institute (R01CA115316, CA115316, R56CA115316)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Caveolin-1 and cellular processes
Keywords
ARF6, tumor invasion, phospholipase D, ERK, myosin light chain kinase, protein kinase C, actomyosin, integrins, extracellular matrix adhesion, protease release, MAPK signaling, tumor progression markers
MeSH terms
ADP-Ribosylation Factor 6, Cell Membrane, Humans, Neoplasm Invasiveness, ADP-Ribosylation Factors

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