Location of Repository

Identifying a role for WASH in the endocytic pathway of Dictyostelium discoideum

By Michael John Carnell

Abstract

Members of the WASP protein family are direct activators of the arp2/3 complex, thereby regulating the nucleation of branched actin assemblies within the cell. Each sub-class possesses a unique N-terminal domain architecture allowing a division of labour between its members, each coupling different signal transduction pathways to the nucleation of specific actin structures. WASH (WASP and SCAR homologue) is a newly identified member of the WASP protein family. Due to its disruption in \(Drosophila\) proving lethal (Linardopopoulou et al., 2007) little is know as to the functional role of WASH at the cellular level. Other than it is important in the development of multicellular organisms. Here we successfully disrupt WASH in the single celled amoebae \(Dictyostelium\) \(discoideum\) and discover a role for WASH in the endocytic pathway. WASH was shown to be essential for the trafficking of indigestible material through the endocytic pathway, with its disruption causing a complete bock in cellular defecation. This was shown to be due to a defect in lysosomal maturation into neutral post-lysosomes. Using fluorescently tagged fusion proteins we show that WASH recruitment coincides with removal of the Vacuolar H+ ATPase from lysosomal membranes, and suggests a possible role for WASH and actin in regulating the luminal pH of intracellular compartments

Topics: QR Microbiology
Year: 2011
OAI identifier: oai:etheses.bham.ac.uk:1550

Suggested articles

Preview

Citations

  1. (2004). A conserved mechanism for Bni1- and mDia1-induced actin assembly and dual regulation of Bni1 by Bud6 and profilin.
  2. (2005). A Dictyostelium homologue of WASP is required for polarized F-actin assembly during chemotaxis.
  3. (2009). A FAM21-containing WASH complex regulates retromer-dependent sorting.
  4. (2000). A myosin I is involved in membrane recycling from early endosomes.
  5. (2008). A novel cellular protein, VPEF, facilitates vaccinia virus penetration into HeLa cells through fluid phase endocytosis.
  6. (2003). A novel role for subunit C in mediating binding of the H+-V-ATPase to the actin cytoskeleton.
  7. (1989). A possible role for Na+,K+-ATPase in regulating ATP-dependent endosome acidification.
  8. (1993). A post-lysosomal compartment in Dictyostelium discoideum.
  9. (2004). A rapid and efficient method to generate multiple gene disruptions in Dictyostelium discoideum using a single selectable marker and the Cre-loxP system.
  10. (1995). A V-ATPase drives active, electrogenic and Na+-independent Cl- absorption across the gills of Eriocheir sinensis.
  11. (1991). A vacuolar-type proton pump energizes K+/H+ antiport in an animal plasma membrane.
  12. (2008). Abi mutants in Dictyostelium reveal specific roles for the SCAR/WAVE complex in cytokinesis. doi
  13. (2003). Abi, Sra1, and Kette control the stability and localization of SCAR/WAVE to regulate the formation of actin-based protrusions.
  14. (2005). Abi1 regulates the activity of N-WASP and WAVE in distinct actin-based processes.
  15. (1984). Acidification of endocytic compartments and the intracellular pathways of ligands and receptors.
  16. (1993). Acidification of rat liver lysosomes: quantitation and comparison with endosomes.
  17. (1986). Acidification of the endocytic and exocytic pathways.
  18. (2006). Actin binding activity of subunit B of vacuolar H+-ATPase is involved in its targeting to ruffled membranes of osteoclasts. doi
  19. (2010). Actin cytoskeleton remodeling by RhoA and RockII regulates vacuolar H+-ATPase (V-ATPase) recycling in epididymal clear cells.
  20. (1995). Actin filament disassembly is a sufficient final trigger for exocytosis in nonexcitable cells.
  21. (2001). Actin pedestal formation by enteropathogenic Escherichia coli and intracellular motility of Shigella flexneri are abolished in N-WASP-defective cells.
  22. (2008). Actin-based propulsive forces and myosin-II-based contractile forces in migrating Dictyostelium cells.
  23. (2000). Actin-dependent propulsion of endosomes and lysosomes by recruitment of N-WASP.
  24. (2002). An actin nucleation mechanism mediated by Bni1 and profilin.
  25. (1994). Analysis of successive endocytic compartments isolated from Dictyostelium discoideum by magnetic fractionation.
  26. (1998). and is essential for endocytosis in yeast.
  27. (2000). ARF1 regulates pH-dependent COP functions in the early endocytic pathway.
  28. (2006). Arp2/3 ATP hydrolysis-catalysed branch dissociation is critical for endocytic force generation.
  29. (2008). Arp2/3 complex activity in filopodia of spreading cells.
  30. (1999). Arp2/3 complex and actin depolymerizing factor/cofilin in dendritic organization and treadmilling of actin filament array in lamellipodia.
  31. (2001). Arp2/3 complex requires hydrolyzable ATP for nucleation of new actin filaments. doi
  32. (2004). ARPC1/Arc40 mediates the interaction of the actin-related protein 2 and 3 complex with Wiskott-Aldrich syndrome protein family activators.
  33. (2003). ATP hydrolysis on actin-related protein 2/3 complex causes debranching of dendritic actin arrays.
  34. (2000). Autoinhibition and activation mechanisms of the Wiskott-Aldrich syndrome protein.
  35. (2008). Biochemical and functional characterization of the actin-binding activity of the B subunit of yeast vacuolar H+-ATPase. doi
  36. (2002). Cadherindirected actin assembly: E-cadherin physically associates with the Arp2/3 complex to direct actin assembly in nascent adhesive contacts.
  37. (1997). cdc12p, a protein required for cytokinesis in fission yeast, is a component of the cell division ring and interacts with profilin.
  38. (1996). Cell motility driven by actin polymerization.
  39. (2003). Cellular motility driven by assembly and disassembly of actin filaments. doi
  40. (2003). Characterization of the WAVE1 knock-out mouse: implications for CNS development.
  41. (1999). Circulation of the plasma membrane in Dictyostelium.
  42. (2002). Conserved interactions with cytoskeletal but not signaling elements are an essential aspect of Drosophila WASp function.
  43. (2006). Control of the assembly of ATP- and ADP-actin by formins and profilin.
  44. (2007). Cordon-bleu is an actin nucleation factor and controls neuronal morphology. doi
  45. (1995). Coronin involved in phagocytosis: dynamics of particle-induced relocalization visualized by a green fluorescent protein Tag.
  46. (2004). Critical conformational changes in the Arp2/3 complex are induced by nucleotide and nucleation promoting factor.
  47. (1967). Cultivation of Dictyostelium discoideum in axenic medium.
  48. (1978). Cytoplasmic streaming in Amoeba proteus is inhibited by the actin-specific drug phalloidin. Exp Cell Res.
  49. (2001). Dendritic organization of actin comet tails.
  50. (2000). Determinants of the pH of the Golgi complex.
  51. (1997). Dictyostelium lysosomal proteins with different sugar modifications sort to functionally distinct compartments.
  52. (1977). Differences in the mode of phagocytosis with Fc and C3 receptors in macrophages.
  53. (1981). Direct measurement of actin polymerization rate constants by electron microscopy of actin filaments nucleated by isolated microvillus cores. doi
  54. (2000). Directed actin polymerization is the driving force for epithelial cell-cell adhesion.
  55. (1995). Disassembly and reassembly of the yeast vacuolar H(+)-ATPase in vivo.
  56. (1989). Disorders of the T-Cell ystem in immunologic disorders in infants and
  57. (1990). Disruption of genes encoding subunits of yeast vacuolar H(+)-ATPase causes conditional lethality.
  58. (2003). Disruption of the Diaphanous-related formin Drf1 gene encoding mDia1 reveals a role for Drf3 as an effector for Cdc42. doi
  59. (2005). Drosophila Spire is an actin nucleation factor. doi
  60. (1967). Effects of cytochalasins on mammalian cells.
  61. (1999). Endocytic vesicles move at the tips of actin tails in cultured mast cells.
  62. (2002). Endosome fusion and microtubulebased dynamics in the early endocytic pathway of dictyostelium. doi
  63. (2002). Essential role of neural Wiskott-Aldrich syndrome protein in podosome formation and degradation of extracellular matrix in src-transformed fibroblasts.
  64. (1988). Establishment of a transient expression system for Dictyostelium discoideum.
  65. (1987). Evidence for ligand- and pHdependent conformational changes in liposome-associated mannose 6-phosphate receptor.
  66. (2003). Evidence for rotation of V1-ATPase.
  67. (2010). Evolutionary conservation of the WASH complex, an actin polymerization machine involved in endosomal fission. doi
  68. (1997). Fluid-phase uptake by macropinocytosis in Dictyostelium.
  69. (2004). Formation of multivesicular endosomes in Dictyostelium.
  70. (2003). Formin leaky cap allows elongation in the presence of tight capping proteins.
  71. (2003). Fusion and fission events in the endocytic pathway of Dictyostelium.
  72. (1974). Genetics of growth in axenic medium of the cellular slime mould Dictyostelium discoideum.
  73. (2000). Growth of Legionella pneumophila in Dictyostelium discoideum: a novel system for genetic analysis of host-pathogen interactions.
  74. (1985). High-copy-number transformants and co-transformation in Dictyostelium.
  75. (2007). Human subtelomeric WASH genes encode a new subclass of the WASP family.
  76. (2006). Identification of a domain in the V0 subunit d that is critical for coupling of the yeast vacuolar proton-translocating ATPase.
  77. (2004). Imaging sites of N-wasp activity in lamellipodia and invadopodia of carcinoma cells.
  78. (1999). Interaction between vacuolar H(+)-ATPase and microfilaments during osteoclast activation.
  79. (2001). Interaction of WASP/Scar proteins with actin and vertebrate
  80. (1996). Interleukin-1 increases vacuolar-type H+-ATPase activity in murine peritoneal macrophages.
  81. (2007). IQGAP1 regulates cell motility by linking growth factor signaling to actin assembly. doi
  82. (2007). IQGAP1 stimulates actin assembly through the N-WASP-Arp2/3 pathway.
  83. (1994). Isolation of a novel gene mutated in Wiskott-Aldrich syndrome.
  84. (1989). Iterative fractionation of recycling receptors from lysosomally destined ligands in an early sorting endosome.
  85. (1993). Kinetics of endosomal pH evolution in Dictyostelium discoideum amoebae. Study by fluorescence spectroscopy. doi
  86. (1992). L. monocytogenes-induced actin assembly requires the actA gene product, a surface protein.
  87. (1983). Latrunculins: novel marine toxins that disrupt microfilament organization in cultured cells.
  88. (2005). leading to actin nucleation.
  89. (2008). Leiomodin is an actin filament nucleator in muscle cells.
  90. (2003). Loss of WAVE-1 causes sensorimotor retardation and reduced learning and memory in mice.
  91. (1983). Low pH accelerates dissociation of receptor-bound insulin.
  92. (2004). Mammalian formin-1 participates in adherens junctions and polymerization of linear actin cables.
  93. Mechanism of acidification of the trans-Golgi network (TGN). In situ measurements of pH using retrieval of TGN38 and furin from the cell surface. doi
  94. (2005). Mechanism of filament nucleation and branch stability revealed by the structure of the Arp2/3 complex at actin branch junctions.
  95. (2003). Mechanism of formin-induced nucleation of actin filaments. doi
  96. (2002). Mechanism of regulation of WAVE1-induced actin nucleation by Rac1 and Nck.
  97. (2001). Membrane sorting in the endocytic and phagocytic pathway of Dictyostelium discoideum.
  98. (1997). Microtubules are involved in bafilomycin A1-induced tubulation and Rab5-dependent vacuolation of early endosomes.
  99. (2005). Modulation of the actin cytoskeleton via gelsolin regulates vacuolar H+-ATPase recycling. doi
  100. (2010). Molecular dissection of Salmonella-induced membrane ruffling versus invasion.
  101. (2005). Molecular mechanisms of invadopodium formation: the role of the N-WASP-Arp2/3 complex pathway and cofilin.
  102. (2003). Molecular requirements for actin-based lamella formation in Drosophila S2 cells.
  103. (2002). Morphology and dynamics of the endocytic pathway in Dictyostelium discoideum.
  104. (1999). Mutations in the gene encoding B1 subunit of H+-ATPase cause renal tubular acidosis with sensorineural deafness.
  105. (2007). Mutations in the KIAA0196 gene at the SPG8 locus cause hereditary spastic paraplegia.
  106. (2004). N-WASP and WAVE2 acting downstream of phosphatidylinositol 3-kinase are required for myogenic cell migration induced by hepatocyte growth factor.
  107. (2005). N-WASP deficiency impairs EGF internalization and actin assembly at clathrin-coated pits. doi
  108. (2001). N-WASP deficiency reveals distinct pathways for cell surface projections and microbial actin-based motility.
  109. (1996). N-WASP, a novel actin-depolymerizing protein, regulates the cortical cytoskeletal rearrangement in a PIP2-dependent manner downstream of tyrosine kinases.
  110. (2006). Nap1 regulates Dictyostelium cell motility and adhesion through SCAR-dependent and -independent pathways.
  111. (2001). Nck and phosphatidylinositol 4,5-bisphosphate synergistically activate actin polymerization through the N-WASP-Arp2/3 pathway.
  112. (2003). Negative regulation of yeast WASp by two SH3 domain-containing proteins.
  113. (1998). Neural Wiskott-Aldrich syndrome protein is implicated in the actin-based motility of Shigella flexneri.
  114. (1998). Noninvasive measurement of the pH of the endoplasmic reticulum at rest and during calcium release.
  115. (2000). Organelle pH studies using targeted avidin and fluoresceinbiotin.
  116. (1989). Osteoclastic bone resorption by a polarized vacuolar proton pump.
  117. (1998). p140Sra-1 (specifically Rac1-associated protein) is a novel specific target for Rac1 small GTPase.
  118. (2009). p53-cofactor JMY is a multifunctional actin nucleation factor.
  119. (2006). Parasexual genetics using axenic cells.
  120. (2002). pH Homeostasis of cellular organelles.
  121. (2001). Phagocytosis and macropinocytosis in Dictyostelium: phosphoinositidebased processes, biochemically distinct.
  122. (2005). Phosphatidylinositol 3-kinase-mediated effects of glucose on vacuolar H+-ATPase assembly, translocation, and acidification of intracellular compartments in renal epithelial cells.
  123. (2003). PIR121 regulates pseudopod dynamics and SCAR activity in Dictyostelium. doi
  124. (1993). Polarized distribution of Listeria monocytogenes surface protein ActA at the site of directional actin assembly.
  125. (2001). Prokaryotic origin of the actin cytoskeleton.
  126. (1989). Proteolytic activity of specialized surface protrusions formed at rosette contact sites of transformed cells.
  127. (1999). Protrusive growth from giant liposomes driven by actin polymerization.
  128. (1994). Purification of a cortical complex containing two unconventional actins from Acanthamoeba by affinity chromatography on profilin-agarose.
  129. (2006). Quantitative and microscopic methods for studying the endocytic pathway.
  130. (1999). Regulation of actin polymerization by Arp2/3 complex and WASp/Scar proteins.
  131. (2008). Regulation of the V-ATPase along the endocytic pathway occurs through reversible subunit association and membrane localization.
  132. (2010). Regulation of vacuolar proton-translocating ATPase activity and assembly by extracellular pH.
  133. (2004). Renal vacuolar H+-ATPase.
  134. (2010). Retrieval of the vacuolar HATPase from phagosomes revealed by live cell imaging. doi
  135. (1995). Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia.
  136. (2002). SCAR is a primary regulator of Arp2/3-dependent morphological events in Drosophila.
  137. (1999). Scar, a WASp-related protein, activates nucleation of actin filaments by the Arp2/3 complex.
  138. (1998). SCAR, a WASP-related protein, isolated as a suppressor of receptor defects in late Dictyostelium development. doi
  139. (1981). Secretion of Lysosomal enzymes in the cellular slime mold, Dictyostelium discoideum.
  140. (1995). Sequences, structural models, and cellular localization of the actin-related proteins
  141. (1997). Structure, subunit topology, and actin-binding activity of the Arp2/3 complex from Acanthamoeba.
  142. (2009). Targeting the actin-binding protein VASP to late endosomes induces the formation of giant actin aggregates.
  143. (2000). The amino-terminal domain of the B subunit of vacuolar H+-ATPase contains a filamentous actin binding site.
  144. (2001). The aminoterminal domain of the vacuolar proton-translocating ATPase a subunit controls targeting and in vivo dissociation, and the carboxyl-terminal domain affects coupling of proton transport and ATP hydrolysis.
  145. (2009). The Arp2/3 activator WASH controls the fission of endosomes through a large multiprotein complex.
  146. (2000). The Arp2/3 complex branches filament barbed ends: functional antagonism with capping proteins.
  147. (1979). The chromaffin granule surface: the presence of actin and the nature of its interaction with the membrane.
  148. (2000). The Dictyostelium RasS protein is required for macropinocytosis, phagocytosis and the control of cell movement.
  149. (2004). The evolution of A-, F-, and V-type ATP synthases and ATPases: reversals in function and changes in the H+/ATP coupling ratio. doi
  150. (2003). The fission yeast cytokinesis formin Cdc12p is a barbed end actin filament capping protein gated by profilin.
  151. (2005). The genome of the social amoeba Dictyostelium discoideum.
  152. (2000). The H subunit (Vma13p) of the yeast VATPase inhibits the ATPase activity of cytosolic V1 complexes. doi
  153. (1998). The interaction of Arp2/3 complex with actin: nucleation, high affinity pointed end capping, and formation of branching networks of filaments.
  154. (2010). The mechanism of rotating proton pumping ATPases. Biochim Biophys Acta.
  155. (2004). The pH of the secretory pathway: measurement, determinants, and regulation. doi
  156. (2000). The recycling endosome of Madin-Darby canine kidney cells is a mildly acidic compartment rich in raft components.
  157. (1992). The role of V-ATPase in neuronal and endocrine systems.
  158. (2008). The structural basis of actin filament branching by the Arp2/3 complex.
  159. (2005). The V-ATPase subunit C binds to polymeric F-actin as well as to monomeric G-actin and induces cross-linking of actin filaments.
  160. (1997). Theoretical considerations on the role of membrane potential in the regulation of endosomal pH.
  161. (2001). Under-agarose folate chemotaxis of Dictyostelium discoideum amoebae in permissive and mechanically inhibited conditions.
  162. (2004). Vacuolar H+-ATPase binding to microfilaments: regulation in response to phosphatidylinositol 3-kinase activity and detailed characterization of the actin-binding site in subunit B.
  163. (1997). VMA11 and VMA16 encode second and third proteolipid subunits of the Saccharomyces cerevisiae vacuolar membrane H+-ATPase.
  164. (2010). WASH and the Arp2/3 complex regulate endosome shape and trafficking.
  165. (2010). WASH and WAVE actin regulators of the Wiskott-Aldrich syndrome protein (WASP) family are controlled by analogous structurally related complexes.
  166. (2009). Wash functions downstream of Rho and links linear and branched actin nucleation factors.
  167. (2010). WASP Family Proteins - Their Evolution and Its Physiological Implications. Mol Biol Cell.
  168. (2001). Wasp, the Drosophila Wiskott-Aldrich syndrome gene homologue, is required for cell fate decisions mediated by Notch signaling. doi
  169. (1998). WAVE, a novel WASP-family protein involved in actin reorganization induced by Rac.
  170. (2003). WAVE2 deficiency reveals distinct roles in embryogenesis and Rac-mediated actinbased motility.
  171. (2003). WAVE2 is required for directed cell migration and cardiovascular development.
  172. (2002). WH2 domain: a small, versatile adapter for actin monomers. doi
  173. (2001). WIP regulates N-WASP-mediated actin polymerization and filopodium formation.
  174. (2005). Wiskott-Aldrich syndrome protein deficiency leads to reduced B-cell adhesion, migration, and homing, and a delayed humoral immune response.
  175. (1998). Wiskott-Aldrich syndrome protein-deficient mice reveal a role for WASP in T but not B cell activation.
  176. (1996). Wiskott-Aldrich syndrome protein, a novel effector for the GTPase CDC42Hs, is implicated in actin polymerization.
  177. (2008). X-ray scattering study of activated Arp2/3 complex with bound actinWCA.

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.