Actin polymerization driven by WASH causes V-ATPase retrieval and vesicle neutralization before exocytosis

Carnell, Michael; Zech, Tobias; Calaminus, Simon D. J.; Ura, Seiji; Hagedorn, Monica; Johnston, Simon A.; May, Robin C.; Soldati, Thierry; Machesky, Laura M.; Insall, Robert H.

doi:10.1083/jcb.201009119

Cited by 143 publications

(195 citation statements)

References 37 publications

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“…1A and Movie S1). Consistent with previous observations of both Dictyostelium postlysosomes and mammalian endosomes (23,24,27), GFP-WASH was restricted to multiple discrete subdomains on the macropinosome surface. This observation indicates a previously unrecognized role for WASH in early macropinosome maturation.…”

Section: Wash Is Recruited To Early Macropinosomes and Phagosomessupporting

confidence: 78%

“…Previously, we showed that a late phase of WASH activity drives the sorting and removal of the V-ATPase during Dictyostelium lysosome to postlysosome transition (27). This finding occurred via direct interaction between the V-ATPase and WASH-generated actin, and arrival of WASH coincides with V-ATPase removal.…”

Section: Wash Is Recruited To Early Macropinosomes and Phagosomesmentioning

confidence: 99%

“…2C). Indeed, phagosomal acidification was actually slightly decreased, most likely due to decreased V-ATPase supply, as so much is sequestered in later compartments such as postlysosomes (27,30 mammalian cells, WASH mediates sorting from endosomes via direct interaction between the FAM21 subunit of the WASH complex and the VPS35 subunit of the retromer sorting complex (24,32,36). The retromer complex drives extraction of specific proteins at several trafficking steps by sequence-specific binding (37)(38)(39).…”

Section: Wash Is Recruited To Early Macropinosomes and Phagosomesmentioning

confidence: 99%

“…In particular, significant progress has been made in defining the role of actin in the segregation and extraction of specific proteins (22). Many studies describe a role for the WASH (WASP and SCAR homolog) complex in driving endosomal actin polymerization (23)(24)(25)(26)(27)(28)(29)(30). WASH has been implicated in the endosome-to-surface trafficking of a number of cargos, including the transferrin receptor (TfnR), α5β1 integrins,…”

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confidence: 99%

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WASH drives early recycling from macropinosomes and phagosomes to maintain surface phagocytic receptors

Buckley

Gopaldass

Bosmani

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Macropinocytosis is an ancient mechanism that allows cells to harvest nutrients from extracellular media, which also allows immune cells to sample antigens from their surroundings. During macropinosome formation, bulk plasma membrane is internalized with all its integral proteins. It is vital for cells to salvage these proteins before degradation, but the mechanisms for sorting them are not known. Here we describe the evolutionarily conserved recruitment of the WASH (WASP and SCAR homolog) complex to both macropinosomes and phagosomes within a minute of internalization. Using Dictyostelium, we demonstrate that WASH drives protein sorting and recycling from macropinosomes and is thus essential to maintain surface receptor levels and sustain phagocytosis. WASH functionally interacts with the retromer complex at both early and late phases of macropinosome maturation, but mediates recycling via retromer-dependent and -independent pathways. WASH mutants consequently have decreased membrane levels of integrins and other surface proteins. This study reveals an important pathway enabling cells to sustain macropinocytosis without bulk degradation of plasma membrane components.phagocytosis | macropinocytosis | WASH | Dictyostelium | trafficking

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Section: Wash Is Recruited To Early Macropinosomes and Phagosomessupporting

confidence: 78%

Section: Wash Is Recruited To Early Macropinosomes and Phagosomesmentioning

confidence: 99%

Section: Wash Is Recruited To Early Macropinosomes and Phagosomesmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

WASH drives early recycling from macropinosomes and phagosomes to maintain surface phagocytic receptors

Buckley

Gopaldass

Bosmani

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…A similar role for WASH in the recycling of integrins has been seen in mammalian cells . Moreover, WASH regulates the retrieval of the vacuolar (V)-ATPase from post-lysosomal compartments, and the recycling of plasma membrane components from early macropinosomes and phagosomes in Dictyostelium (Buckley et al, 2016;Carnell et al, 2011). Thus, the ability of WASH to regulate receptor recycling and lysosome neutralization has been highly conserved throughout evolution.…”

Section: Jmymentioning

confidence: 99%

Cellular functions of WASP family proteins at a glance

Alekhina

Burstein

Billadeau

2017

Journal of Cell Science

158

172

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Proteins of the Wiskott–Aldrich syndrome protein (WASP) family function as nucleation-promoting factors for the ubiquitously expressed Arp2/3 complex, which drives the generation of branched actin filaments. Arp2/3-generated actin regulates diverse cellular processes, including the formation of lamellipodia and filopodia, endocytosis and/or phagocytosis at the plasma membrane, and the generation of cargo-laden vesicles from organelles including the Golgi, endoplasmic reticulum (ER) and the endo-lysosomal network. Recent studies have also identified roles for WASP family members in promoting actin dynamics at the centrosome, influencing nuclear shape and membrane remodeling events leading to the generation of autophagosomes. Interestingly, several WASP family members have also been observed in the nucleus where they directly influence gene expression by serving as molecular platforms for the assembly of epigenetic and transcriptional machinery. In this Cell Science at a Glance article and accompanying poster, we provide an update on the subcellular roles of WHAMM, JMY and WASH (also known as WASHC1), as well as their mechanisms of regulation and emerging functions within the cell.

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Copine A regulates the size and exocytosis of contractile vacuoles and postlysosomes in Dictyostelium

2020

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Copines are a family of cytosolic proteins that associate with membranes in a calcium‐dependent manner and are found in many eukaryotic organisms. Dictyostelium discoideum has six copine genes (cpnA‐cpnF), and cells lacking cpnA(cpnA−) have defects in cytokinesis, chemotaxis, adhesion, and development. CpnA has also been shown to associate with the plasma membrane, contractile vacuoles (CV), and organelles of the endolysosomal pathway. Here, we use cpnA− cells to investigate the role of CpnA in CV function and endocytosis. When placed in water, cpnA− cells made abnormally large CVs that took longer to expel. Visualization of CVs with the marker protein GFP‐dajumin indicated that cpnA− cells had fewer CVs that sometimes refilled before complete emptying. In endocytosis assays, cpnA− cells took up small fluorescent beads by macropinocytosis at rates similar to parental cells. However, cpnA− cells reached a plateau sooner than parental cells and had less fluorescence at later time points. p80 antibody labeling of postlysosomes (PL) indicated that there were fewer and smaller PLs in cpnA− cells. In dextran pulse‐chase experiments, the number of PLs peaked earlier in cpnA− cells, and the PLs did not become as large and disappeared sooner as compared to parental cells. PLs in cpnA− cells were also shown to have more actin coats, suggesting CpnA may play a role in actin filament disassembly on PL membranes. Overall, these results indicate that CpnA is involved in the regulation of CV size and expulsion, and the maturation, size, and exocytosis of PLs.

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Actin polymerization driven by WASH causes V-ATPase retrieval and vesicle neutralization before exocytosis

Abstract: WASH coats mature lysosomes and is required for exocytosis of indigestible material.

Cited by 143 publications

References 37 publications

WASH drives early recycling from macropinosomes and phagosomes to maintain surface phagocytic receptors

WASH drives early recycling from macropinosomes and phagosomes to maintain surface phagocytic receptors

Cellular functions of WASP family proteins at a glance

Copine A regulates the size and exocytosis of contractile vacuoles and postlysosomes in Dictyostelium

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