[en] The lectin isolated from Xerocomus chrysenteron (XCL) displays a toxic activity towards insects. In order to assess its possible mode of action and to gather useful data for its potential use in insect-resistant transgenic plants, we investigated the effects of XCL at the cellular level. Immunofluorescence microscopy studies revealed that XCL is rapidly internalized into small endocytic vesicles that further coalesce in the perinuclear region. We show that XCL is endocytosed by the clathrin-dependent pathway, and is delivered to late endosome/lysosome compartments. The internalization of XCL seems to be general since it occurs in different cell types such as insect (SF9) or mammalian (NIH-3T3 and Hela) cell lines. In the presence of XCL, the uptake of GFP and BSA is greatly enhanced, demonstrating that XCL facilitates endocytosis. Thus, XCL could serve as a delivery agent to facilitate the endocytosis of proteins that do not enter the cell alone.
Aubin, J. E., Tolson, N., Ling, V. (1980): The redistribution of fluoresceinated concanavalin A in Chinese hamster ovary cells and in their colcemid-resistant mutants. Exp. Cell Res. 126, 75-85.
Balin, B. J., Broadwell, R. D. (1987): Lectin-labeled membrane is transferred to the Golgi complex in mouse pituitary cells in vivo. J. Histochem. Cytochem. 35, 489-498.
Batra, R. K., Wang-Johanning, F., Wagner, E., Garver, R. I., Jr., Curiel, D. T. (1994): Receptor mediated gene delivery employing lectin-binding specificity. Gene Ther. 1, 255-260.
Benmerah, A., Lamaze, C. (2002): Endocytose: Chaque voie compte! Medecine/Sciences 18, 1126-1136
Conner, S. D., Schmid, S. L. (2003): Regulated portals of entry into the cell. Nature 422, 37-44.
Falnes, P. O., Sandvig, K. (2000): Penetration of protein toxins into cells. Curr. Opin. Cell Biol. 12, 407-413.
Gatehouse, A. M. R., Down, R. E., Powell, K. S., Sauvion, N., Rahbé, Y., Newell, C. A., Merryweather, A., Hamilton, W. D. O., Gatehouse, J. A. (1996): Transgenic potato plants with enhanced resistance to peach-potato aphid Myzus persicae. Entomol. Exp. Applic. 79, 295-307.
Goldstein, I. J., Hughes, R. C., Monsigny, M., Osawa, T., Sharon, N. (1980): What should be called a lectin. Nature 285, 66-67.
Guillot, J., Konska, G. (1997): Lectins in higher fungi. Biochem. System. Ecol. 25, 203-230.
Harper, S. M., Crenshaw, R. W., Mullins, M. A., Privalle L. S. (1995): Lectin binding to insect brush border membranes. J. Econ. Entomol. 88, 1197-1202.
Ippoliti, R., Lendaro, E., Benedetti, P. A., Torrisi, M. R., Belleudi, F., Carpani, D., Soria, M. R., Fabbrini, M. S. (2000): Endocytosis of a chimera between human pro-urokinase and the plant toxin saporin: an unusual internalization mechanism. FASE9 J. 14, 1335-1344.
Janzen, D. H., Juster, H. B., Liener, I. R. (1976): Insecticidal action of the phytohemagglutinin in black beans on a Bruchid beetle. Science 192, 795-796.
Kirchhausen, T. (2000): Three ways to make a vesicle. Nat. Rev. Mol. Cell Biol. 1, 187-198.
Lamazel, C., Dujeancourt, A., Baba, T., Lo, C. G., Benmerah, A., Dautry-Varsat A. (2001): Interleukin 2 receptors and detergent-resistant membrane domains define a clathrin-independent endocytic pathway. Mol. Cell 7, 661-671.
Larkin, J. M., Brown, M. S., Goldstein J. L., Anderson, R. G. (1983): Depletion of intracellular potassium arrests coated pit formation and receptor-mediated endocytosis in fibroblasts. Cell 33, 273-285.
Ludwig, T., Griffiths, G., Hoflack, B. (1991): Distribution of newly synthesized lysosomal enzymes in the endocytic pathway of normal rat kidney cells. J Cell Biol. 115, 1561-72.
Mallard, F., Antony, C., Tenza, D., Salamero, J., Goud, B., Johannes, L. (1998): Direct pathway from early/recycling endosomes to the Golgi apparatus revealed through the study of Shiga toxin B-fragment transport. J. Cell Biol. 143, 973-990.
Mier, N., Canete, S., Klaebe, A., Chavant, L., Fournier, D. (1996): Insecticidal properties of mushroom and toadstool carpophores. Phytochemistry 41, 1293-1299.
Mody, R., Joshi, S., Chaney, W. (1995): Use of lectins as diagnostic and therapeutic tools for cancer. J. Pharmacol. Toxicol. Methods 33, 1-10.
Olsnes, S., Kozlov, J. V. (2001): Ricin. Toxicon 39, 1723-1728.
Orlandi, P. A., Fishman, P. H. (1998) Filipin-dependent inhibition of cholera toxin: evidence for toxin internalization and activation through caveolae-like domains. J. Cell Biol. 141, 905-915.
Osborn, T. C., Alexander, D. C., Sun, S. S. M., Cardona, C., Bliss, F. A. (1988): Insecticidal activity and lectin homology of arcelin seed protein. Science 240, 207-210.
Pavelka, M., Ellinger, A., Debbage, P., Loewe, C., Vetterlein, M., Roth, J. (1998): Endocytic routes to the Golgi apparatus. Histochem. Cell Biol. 109, 555-570.
Pelkmans, L., Kartenbeck, J., Helenius, A. (2001): Caveolar endocytosis of simian virus 40 reveals a new two-step vesicular transport pathway to the ER. Nat. Cell Biol. 3, 473-483.
Peumans, W. J., Van Damme, J. M. (1995). Lectins as plant defense proteins. Plant Physiol. 109, 347-352.
Powell, K. S., Spence, J., Bharathi, M., Gatehouse, J. A., Gatehouse, A. M. R. (1998): Immunohistochemicals and developmental studies to elucidate the mechanism of action of the snowdrop lectin on the rice brown plant hopper, Nilaparvata lugens. J. Insect Physiol. 44, 529-539.
Rosén, S., Ek, B., Rask, L., Tunlid, A. (1992): Purification and characterization of a surface lectin from the nematode-trapping fungus Arthrobotrys of oligospora. J. Gen. Microbiol. 138, 2663-2672.
Sandvig, K., Olsnes, S., Brown, J. E., Petersen, O. W., van Deurs, B. (1989): Endocytosis from coated pits of Shiga toxin: a glycolipid-binding protein from Shigella dysenteriae 1. J. Cell Biol. 108, 1331-1343.
Sandvig, K., van Deurs, B. (2000): Entry of ricin and Shiga toxin into cells: molecular mechanisms and medical perspectives. EMBO J. 19, 5943-5950.
Sandvig, K., Grimmer, S., Lauvrak, S. U., Torgersen, M. L., Skretting, G., van Deurs, B., Iversen, T. G. (2002): Pathways followed by ricin and Shiga toxin into cells. Histochem. Cell Biol. 117, 131-141.
Sauvion, N., Rahbé, Y., Peumans, W. J., Van Damme, E. J. M., Gatehouse, J. A., Gatehouse, M. R. (1996): Effects of GNA and other mannose binding lectins on development and fecundity of the peach-potato aphid Myzus persicae. Entomol. Exp. Applic. 79, 285-293.
Schwarz, M. A., Harper, P. A., Juliano, R. L. (1982): Interactions of lectins with CHO cell surface membranes. II. Differential effects of local anaesthetics on endocytosis of Con A and WGA binding sites. J. Cell. Physiol. 111, 264-274.
Schnitzer, J. E., Oh, P., McIntosh, D. P. (1996): Role of GTP hydrolysis in fission of caveolae directly from plasma membranes. Science 274, 239-242.
Shogomori, H., Futerman, A. H. (2001): Cholera toxin is found in detergent-insoluble rafts/domains at the cell surface of hippocampal neurons but is internalized via a raft-independent mechanism. J. Biol. Chem. 276, 9182-9188.
Simons, K., Gruenberg, J. (2000): Jamming the endosomal system: lipid rafts and lysosomal storage diseases. Trends Cell Biol. 10, 459-462.
Sorkin, A. (2000): The endocytosis machinery. J. Cell Sci. 113, 4375-4376.
Subtil, A., Hémar, A., Dautry-Varsat, A. (1994): Rapid endocytosis of interleukin 2 receptors when clathrin-coated pit endocytosis is inhibited. J. Cell Sci. 107, 3461-3468.
Suzaki, E., Kataoka, K. (1999): Three-dimensional visualization of the Golgi apparatus: observation of Brunner's gland cells by a confocal laser scanning microscope. J. Struct. Biol. 128, 131-138.
Torgersen, M. L., Skretting, G., van Deurs, B., Sandvig, K. (2001): Internalization of cholera toxin by different endocytic mechanisms. J. Cell Sci. 114, 3737-3747.
Trigueros, V., Wang, M., Pere, D., Paquereau, L., Chavant, L., Fournier, D. (2000): Modulation of a lectin insecticidal activity by carbohydrates. Arch. Insect Biochem. Physiol. 45, 175-179.
Trigueros, V., Lougarre, A., Ali-Ahmed, D., Rabhe, Y., Guillot, J., Chavant, L., Fournier, D., Paquereau L. (2003): Xerocomus chrysenteron lectin: identification of a new pesticidal protein. Biochim. Biophys. Acta 1621, 292-298.
van Deurs, B., Petersen, O. W., Olsnes, S., Sandvig, K. (1989): The ways of endocytosis. Int. Rev. Cytol. 117, 131-77.
Vetterlein, M., Ellinger, A., Neumüller J., Pavelka, M. (2002): Golgi apparatus and TGN during endocytosis. Histochem. Cell Biol. 117, 143-150.
von Figura, K., Hasilik, A. (1986): Lysosomal enzymes and their receptors. Annu. Rev. Biochem. 55, 167-193.
Wang, L. H., Rothberg, K. G., Anderson, R. G. (1993): Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. J. Cell Biol. 123, 1107-1117.
Wang, M., Trigueros, V., Paquereau, L., Chavant, L., Fournier D. (2002): Proteins as active compounds involved in insecticidal activity of mushroom fruitbodies. J. Econ. Entomol. 95, 603-607.
Wu, C. H., Yeh, S. T., Ling, E. A. (1999): Studies of lectin receptors of rat microglia in culture: receptor distribution and internalization. Exp. Brain Res. 124, 89-99.
Yanagihara, K., Cheng, P. W. (1999): Lectin enhancement of the lipofection efficiency in human lung carcinoma cells. Biochim. Biophys. Acta 1472, 25-33.
Yin, W., Cheng, P. W. (1994): Lectin conjugate-directed gene transfer to airway epithelial cells. Biochem. Biophys. Res. Commun. 205, 826-833.
Yu, L. G., Fernig, D. G., White, M. R., Spiller, D. G., Appleton, P., Evans, R. C., Grierson, I., Smith, J. A., Davies, H., Gerasimenko, O. V., Petersen, O. H., Milton, J. D., Rhodes, J. M. (1999): Edible mushroom (Agaricus bisporus) lectin, which reversibly inhibits epithelial cell proliferation, blocks nuclear localization sequence-dependent nuclear protein import. J. Biol. Chem. 274, 4890-4899.
