Macropinocytosis: regulated coordination of endocytic and exocytic membrane traffic events
10.1242/jcs.03238
Crossref journal-article
The Company of Biologists
Journal of Cell Science (237)
Abstract

Macropinocytosis, a form of bulk uptake of fluid and solid cargo into cytoplasmic vacuoles, called macropinosomes, has been studied mostly in relation to antigen presentation. Early membrane traffic events occurring in this process are, however, largely unknown. Using human dendritic cells we show that a marked increase in the rate of macropinocytosis occurs a few minutes after application of two markers (small latex beads or dextran), depends on a slow intracellular Ca2+ concentration ([Ca2+]i) rise that precedes the PI3K-dependent step, and is preceded and accompanied by exocytosis of enlargeosomes compensating in part for the macropinocytic plasma membrane internalization. Unexpectedly, macropinosomes themselves, which share markers with endosomes, undergo Ca2+-dependent exocytosis so that, after ∼20 minutes of continuous bead or dextran uptake, an equilibrium is reached preventing cells from overloading themselves with the organelles. Large [Ca2+]i increases induced by ionomycin trigger rapid (<1 minute) exocytic regurgitation of all macropinosomes, whereas endosomes remain apparently unaffected. We conclude that, in dendritic cells, the rate of macropinocytosis is not constant but increases in a regulated fashion, as previously shown in other cell types. Moreover, macropinosomes are not simple containers that funnel cargo to an endocytic pathway, but unique organelles, distinct from endosomes by their competence for regulated exocytosis and other membrane properties.

Bibliography

Falcone, S., Cocucci, E., Podini, P., Kirchhausen, T., Clementi, E., & Meldolesi, J. (2006). Macropinocytosis: regulated coordination of endocytic and exocytic membrane traffic events. Journal of Cell Science, 119(22), 4758–4769.

Authors 6
  1. Sestina Falcone (first)
  2. Emanuele Cocucci (additional)
  3. Paola Podini (additional)
  4. Tomas Kirchhausen (additional)
  5. Emilio Clementi (additional)
  6. Jacopo Meldolesi (additional)
References 57 Referenced 217
  1. Amyere, M., Payrastre, B., Krause, U., Van Der Smissen, P., Veithen, A. and Courtoy, P. J. (2000). Constitutive macropinocytosis in oncogene-transformed fibroblasts depends on sequential permanent activation of phosphoinositide 3-kinase and phospholipase C. Mol. Biol. Cell11, 3453-3467. (10.1091/mbc.11.10.3453)
  2. Amyere, M., Mettlen, M., Van Der Smissen, P., Platek, A., Payrastre, B., Veithen, A. and Courtoy, P. J. (2002). Origin, originality, functions, subversions and molecular signalling of macropinocytosis. Int. J. Med. Microbiol.291, 487-494.
  3. Araki, N., Johnson, M. T. and Swanson, J. A. (1996). A role for phosphoinositide 3-kinase in the completion of macropinocytosis and phagocytosis by macrophages. J. Cell Biol.135, 1249-1260. (10.1083/jcb.135.5.1249)
  4. Araki, N., Hatae, T., Furukawa, A. and Swanson, J. A. (2003). Phosphoinositide-3-kinase-independent contractile activities associated with Fcgamma-receptor-mediated phagocytosis and macropinocytosis in macrophages. J. Cell Sci.116, 247-257. (10.1242/jcs.00235)
  5. Bajno, L., Peng, X. R., Schreiber, A. D., Moore, H. P., Trimble, W. S. and Grinstein, S. (2000). Focal exocytosis of VAMP3-containing vesicles at sites of phagosome formation. J. Cell Biol.149, 697-706. (10.1083/jcb.149.3.697)
  6. Borgonovo, B., Cocucci, E., Racchetti, G., Podini, P., Bachi, A. and Meldolesi, J. (2002). Regulated exocytosis: a novel, widely expressed system. Nat. Cell Biol.4, 955-962. (10.1038/ncb888)
  7. Cerny, J., Feng, Y., Yu, A., Miyake, K., Borgonovo, B., Klumperman, J., Meldolesi, J., McNeil, P. L. and Kirchhausen, T. (2004). The small chemical vacuolin-1 inhibits Ca(2+)-dependent lysosomal exocytosis but not cell resealing. EMBO Rep.5, 883-888. (10.1038/sj.embor.7400243)
  8. Cerny, J., Feng, Y., Yu, A., Miyake, K., Borgonovo, B., Klumperman, J., Meldolesi, J., McNeil, P. L. and Kirchhausen, T. (2005). Corrigendum: The small chemical vacuolin-1 inhibits Ca(2+)-dependent lysosomal exocytosis but not cell resealing. EMBO Rep.9, 898.
  9. Cochilla, A. J., Angleson, J. K. and Betz, W. J. (1999). Monitoring secretory membrane with FM1-43 fluorescence. Annu. Rev. Neurosci.22, 1-10. (10.1146/annurev.neuro.22.1.1)
  10. Cocucci, E., Racchetti, G., Podini, P., Rupnik, M. and Meldolesi, J. (2004). Enlargeosome, an exocytic vesicle resistant to nonionic detergents, undergoes endocytosis via a nonacidic route. Mol. Biol. Cell15, 5356-5368. (10.1091/mbc.e04-07-0577)
  11. de Baey, A. and Lanzavecchia, A. (2000). The role of aquaporins in dendritic cell macropinocytosis. J. Exp. Med.191, 743-748. (10.1084/jem.191.4.743)
  12. Dharmawardhane, S., Schurmann, A., Sells, M. A., Chernoff, J., Schmid, S. L. and Bokoch, G. M. (2000). Regulation of macropinocytosis by p21-activated kinase-1. Mol. Biol. Cell11, 3341-3352. (10.1091/mbc.11.10.3341)
  13. Di, A., Nelson, D. J., Bindokas, V., Brown, M. E., Libunao, F. and Palfrey, H. C. (2003). Dynamin regulates focal exocytosis in phagocytosing macrophages. Mol. Biol. Cell14, 2016-2028. (10.1091/mbc.e02-09-0626)
  14. Ellerbroek, S. M., Wennerberg, K., Arthur, W. T., Dunty, J. M., Bowman, D. R., DeMali, K. A., Der, C. and Burridge, K. (2004). SGEF, a RhoG guanine nucleotide exchange factor that stimulates macropinocytosis. Mol. Biol. Cell15, 3309-3319. (10.1091/mbc.e04-02-0146)
  15. Fiorentini, C., Falzano, L., Fabbri, A., Stringaro, A., Logozzi, M., Travaglione, S., Contamin, S., Arancia, G., Malorni, W. and Fais, S. (2001). Activation of rho GTPases by cytotoxic necrotizing factor 1 induces macropinocytosis and scavenging activity in epithelial cells. Mol. Biol. Cell12, 2061-2073. (10.1091/mbc.12.7.2061)
  16. Garrett, W. S., Chen, L. M., Kroschewski, R., Ebersold, M., Turley, S., Trombetta, S., Galan, J. E. and Mellman, I. (2000). Developmental control of endocytosis in dendritic cells by Cdc42. Cell102, 325-334. (10.1016/S0092-8674(00)00038-6)
  17. Gekle, M., Freudinger, R. and Mildenberger, S. (2001). Inhibition of Na+-H+ exchanger-3 interferes with apical receptor-mediated endocytosis via vesicle fusion. J. Physiol.531, 619-629. (10.1111/j.1469-7793.2001.0619h.x)
  18. Hamasaki, M., Araki, N. and Hatae, T. (2004). Association of early endosomal autoantigen 1 with macropinocytosis in EGF-stimulated A431 cells. Anat. Rec. A Discov. Mol. Cell Evol. Biol.277, 298-306. (10.1002/ar.a.20027)
  19. Harding, C. V. and Geuze, H. J. (1992). Class II MHC molecules are present in macrophage lysosomes and phagolysosomes that function in the phagocytic processing of Listeria monocytogenes for presentation to T cells. J. Cell Biol.119, 531-542. (10.1083/jcb.119.3.531)
  20. Henson, P. M., Bratton, D. L. and Fadok, V. A. (2001). Apoptotic cell removal. Curr. Biol.11, R795-R805. (10.1016/S0960-9822(01)00474-2)
  21. Hewlett, L. J., Prescott, A. R. and Watts, C. (1994). The coated pit and macropinocytic pathways serve distinct endosome populations. J. Cell Biol.124, 689-703. (10.1083/jcb.124.5.689)
  22. Huynh, C. and Andrews, N. W. (2005). The small chemical vacuolin-1 alters the morphology of lysosomes without inhibiting Ca2+-regulated exocytosis. EMBO Rep.6, 843-847. (10.1038/sj.embor.7400495)
  23. Knight, D. E. (2002). Calcium-dependent transferrin receptor recycling in bovine chromaffin cells. Traffic3, 298-307. (10.1034/j.1600-0854.2002.030407.x)
  24. Kolb-Maurer, A., Wilhelm, M., Weissinger, F., Brocker, E. B. and Goebel, W. (2002). Interaction of human hematopoietic stem cells with bacterial pathogens. Blood100, 3703-3709. (10.1182/blood-2002-03-0898)
  25. Kruth, H. S., Jones, N. L., Huang, W., Zhao, B., Ishii, I., Chang, J., Combs, C. A., Malide, D. and Zhang, W. Y. (2005). Macropinocytosis is the endocytic pathway that mediates macrophage foam cell formation with native low density lipoprotein. J. Biol. Chem.280, 2352-2360. (10.1074/jbc.M407167200)
  26. Lanzetti, L., Palamidessi, A., Areces, L., Scita, G. and Di Fiore, P. P. (2004). Rab5 is a signalling GTPase involved in actin remodelling by receptor tyrosine kinases. Nature429, 309-314. (10.1038/nature02542)
  27. Lindmo, K. and Stenmark, H. (2006). Regulation of membrane traffic by phosphoinositide 3-kinases. J. Cell Sci.119, 605-614. (10.1242/jcs.02855)
  28. Lutz, M. B., Rovere, P., Kleijmeer, M. J., Rescigno, M., Assmann, C. U., Oorschot, V. M., Geuze, H. J., Trucy, J., Demandolx, D., Davoust, J. et al. (1997). Intracellular routes and selective retention of antigens in mildly acidic cathepsin D/lysosome-associated membrane protein-1/MHC class II-positive vesicles in immature dendritic cells. J. Immunol.159, 3707-3716. (10.4049/jimmunol.159.8.3707)
  29. Maxfield, F. R. and McGraw, T. E. (2004). Endocytic recycling. Nat. Rev. Mol. Cell Biol.5, 121-132. (10.1038/nrm1315)
  30. Meier, O. and Greber, U. F. (2004). Adenovirus endocytosis. J. Gene Med.6, S152-S163. (10.1002/jgm.553)
  31. Meier, O., Boucke, K., Hammer, S. V., Keller, S., Stidwill, R. P., Hemmi, S. and Greber, U. F. (2002). Adenovirus triggers macropinocytosis and endosomal leakage together with its clathrin-mediated uptake. J. Cell Biol.158, 1119-1131. (10.1083/jcb.200112067)
  32. Mellman, I. and Steinman, R. M. (2001). Dendritic cells: specialized and regulated antigen processing machines. Cell106, 255-258. (10.1016/S0092-8674(01)00449-4)
  33. Morris, C. E. and Homann, U. (2001). Cell surface area regulation and membrane tension. J. Membr. Biol.179, 79-102. (10.1007/s002320010040)
  34. Muro, S., Wiewrodt, R., Thomas, A., Koniaris, L., Albelda, S. M., Muzykantov, V. R. and Koval, M. (2003). A novel endocytic pathway induced by clustering endothelial ICAM-1 or PECAM-1. J. Cell Sci.116, 1599-1609. (10.1242/jcs.00367)
  35. Ozaki, S., DeWald, D. B., Shope, J. C., Chen, J. and Prestwich, G. D. (2000). Intracellular delivery of phosphoinositides and inositol phosphates using polyamine carriers. Proc. Natl. Acad. Sci. USA97, 11286-11291. (10.1073/pnas.210197897)
  36. Paolucci, C., Rovere, P., De Nadai, C., Manfredi, A. A. and Clementi, E. (2000). Nitric oxide inhibits the tumor necrosis factor alpha-regulated endocytosis of human dendritic cells in a cyclic GMP-dependent way. J. Biol. Chem.275, 19638-19644. (10.1074/jbc.M000511200)
  37. Paolucci, C., Burastero, S. E., Rovere-Querini, P., De Palma, C., Falcone, S., Perrotta, C., Capobianco, A., Manfredi, A. A. and Clementi, E. (2003). Synergism of nitric oxide and maturation signals on human dendritic cells occurs through a cyclic GMP-dependent pathway. J. Leukoc. Biol.73, 253-262. (10.1189/jlb.0902447)
  38. Pelkmans, L. and Helenius, A. (2003). Insider information: what viruses tell us about endocytosis. Curr. Opin. Cell Biol.15, 414-422. (10.1016/S0955-0674(03)00081-4)
  39. Racoosin, E. L. and Swanson, J. A. (1993). Macropinosome maturation and fusion with tubular lysosomes in macrophages. J. Cell Biol.121, 1011-1020. (10.1083/jcb.121.5.1011)
  40. Reece, J. C., Vardaxis, N. J., Marshall, J. A., Crowe, S. M. and Cameron, P. U. (2001). Uptake of HIV and latex particles by fresh and cultured dendritic cells and monocytes. Immunol. Cell Biol.79, 255-263. (10.1046/j.1440-1711.2001.01011.x)
  41. Rupper, A., Lee, K., Knecht, D. and Cardelli, J. (2001). Sequential activities of phosphoinositide 3-kinase, PKB/Akt, and Rab7 during macropinosome formation in Dictyostelium. Mol. Biol. Cell12, 2813-2824. (10.1091/mbc.12.9.2813)
  42. Sallusto, F., Cella, M., Danieli, C. and Lanzavecchia, A. (1995). Dendritic cells use macropinocytosis and the mannose receptor to concentrate macromolecules in the major histocompatibility complex class II compartment: downregulation by cytokines and bacterial products. J. Exp. Med.182, 389-400. (10.1084/jem.182.2.389)
  43. Salter, R. D., Tuma-Warrino, R. J., Hu, P. Q. and Watkins, S. C. (2004). Rapid and extensive membrane reorganization by dendritic cells following exposure to bacteria revealed by high-resolution imaging. J. Leukoc. Biol.75, 240-243. (10.1189/jlb.0703339)
  44. Schnatwinkel, C., Christoforidis, S., Lindsay, M. R., Uttenweiler-Joseph, S., Wilm, M., Parton, R. G. and Zerial, M. (2004). The Rab5 effector Rabankyrin-5 regulates and coordinates different endocytic mechanisms. PLoS Biol.2, E261. (10.1371/journal.pbio.0020261)
  45. Sheff, D. R., Daro, E. A., Hull, M. and Mellman, I. (1999). The receptor recycling pathway contains two distinct populations of early endosomes with different sorting functions. J. Cell Biol.145, 123-139. (10.1083/jcb.145.1.123)
  46. Steinman, R. M. and Swanson, J. (1995). The endocytic activity of dendritic cells. J. Exp. Med.182, 283-288. (10.1084/jem.182.2.283)
  47. Sun, P., Yamamoto, H., Suetsugu, S., Miki, H., Takenawa, T. and Endo, T. (2003). Small GTPase Rah/Rab34 is associated with membrane ruffles and macropinosomes and promotes macropinosome formation. J. Biol. Chem.278, 4063-4071. (10.1074/jbc.M208699200)
  48. Swanson, J. A. and Watts, C. (1995). Macropinocytosis. Trends Cell Biol.5, 424-428. (10.1016/S0962-8924(00)89101-1)
  49. Swanson, J. A., Johnson, M. T., Beningo, K., Post, P., Mooseker, M. and Araki, N. (1999). A contractile activity that closes phagosomes in macrophages. J. Cell Sci.112, 307-316. (10.1242/jcs.112.3.307)
  50. Terebiznik, M. R., Vieira, O. V., Marcus, S. L., Slade, A., Yip, C. M., Trimble, W. S., Meyer, T., Finlay, B. B. and Grinstein, S. (2002). Elimination of host cell PtdIns(4,5)P(2) by bacterial SigD promotes membrane fission during invasion by Salmonella. Nat. Cell Biol.4, 766-773. (10.1038/ncb854)
  51. van der Wijk, T., Tomassen, S. F., Houtsmuller, A. B., de Jonge, H. R. and Tilly, B. C. (2003). Increased vesicle recycling in response to osmotic cell swelling. Cause and consequence of hypotonicity-provoked ATP release. J. Biol. Chem.278, 40020-40025. (10.1074/jbc.M307603200)
  52. Veithen, A., Amyere, M., Van Der Smissen, P., Cupers, P. and Courtoy, P. J. (1998). Regulation of macropinocytosis in v-Src-transformed fibroblasts: cyclic AMP selectively promotes regurgitation of macropinosomes. J. Cell Sci.111, 2329-2335. (10.1242/jcs.111.16.2329)
  53. Wadia, J. S., Stan, R. V. and Dowdy, S. F. (2004). Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis. Nat. Med.10, 310-315. (10.1038/nm996)
  54. West, M. A., Bretscher, M. S. and Watts, C. (1989). Distinct endocytotic pathways in epidermal growth factor-stimulated human carcinoma A431 cells. J. Cell Biol.109, 2731-2739. (10.1083/jcb.109.6.2731)
  55. West, M. A., Prescott, A. R., Eskelinen, E. L., Ridley, A. J. and Watts, C. (2000). Rac is required for constitutive macropinocytosis by dendritic cells but does not control its downregulation. Curr. Biol.10, 839-848. (10.1016/S0960-9822(00)00595-9)
  56. West, M. A., Wallin, R. P., Matthews, S. P., Svensson, H. G., Zaru, R., Ljunggren, H. G., Prescott, A. R. and Watts, C. (2004). Enhanced dendritic cell antigen capture via toll-like receptor-induced actin remodeling. Science305, 1153-1157. (10.1126/science.1099153)
  57. Yang, Z., Vadlamudi, R. K. and Kumar, R. (2005). Dynein light chain 1 phosphorylation controls macropinocytosis. J. Biol. Chem.280, 654-659. (10.1074/jbc.M408486200)
Dates
Type When
Created 18 years, 10 months ago (Oct. 31, 2006, 11:18 p.m.)
Deposited 2 years, 3 months ago (May 9, 2023, 11:55 a.m.)
Indexed 5 days, 6 hours ago (Aug. 27, 2025, 12:35 p.m.)
Issued 18 years, 9 months ago (Nov. 15, 2006)
Published 18 years, 9 months ago (Nov. 15, 2006)
Published Print 18 years, 9 months ago (Nov. 15, 2006)
Funders 0

None

@article{Falcone_2006, title={Macropinocytosis: regulated coordination of endocytic and exocytic membrane traffic events}, volume={119}, ISSN={0021-9533}, url={http://dx.doi.org/10.1242/jcs.03238}, DOI={10.1242/jcs.03238}, number={22}, journal={Journal of Cell Science}, publisher={The Company of Biologists}, author={Falcone, Sestina and Cocucci, Emanuele and Podini, Paola and Kirchhausen, Tomas and Clementi, Emilio and Meldolesi, Jacopo}, year={2006}, month=nov, pages={4758–4769} }