Abstract
We have previously demonstrated that the preendosomal compartment in addition to clathrin-coated vesicles, comprises distinct nonclathrin coated endocytic vesicles mediating clathrin-independent endocytosis (Hansen, S. H., K. Sandvig, and B. van Deurs. 1991. J. Cell Biol. 113:731-741). Using K+ depletion in HEp-2 cells to block clathrin- dependent but not clathrin-independent endocytosis, we have now traced the intracellular routing of these nonclathrin coated vesicles to see whether molecules internalized by clathrin-independent endocytosis are delivered to a unique compartment or whether they reach the same early and late endosomes as encountered by molecules internalized with high efficiency through clathrin-coated pits and vesicles. We find that Con A-gold internalized by clathrin-independent endocytosis is delivered to endosomes containing transferrin receptors. After incubation of K(+)- depleted cells with Con A-gold for 15 min, approximately 75% of Con A- gold in endosomes is colocalized with transferrin receptors. Endosomes containing only Con A-gold may be accounted for either by depletion of existing endosomes for transferrin receptors or by de novo generation of endosomes. Cationized gold and BSA-gold internalized in K(+)- depleted cells are also delivered to endosomes containing transferrin receptors. h-lamp-1-enriched compartments are only reached occasionally within 30 min in K(+)-depleted as well as in control cells. Thus, preendosomal vesicles generated by clathrin-independent endocytosis do not fuse to any marked degree with late endocytic compartments. These data show that in HEp-2 cells, molecules endocytosed without clathrin are delivered to the same endosomes as reached by transferrin receptors internalized through clathrin-coated pits.
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- Bamezai A., Goldmacher V. S., Rock K. L. Internalization of glycosyl-phosphatidylinositol (GPI)-anchored lymphocyte proteins. II. GPI-anchored and transmembrane molecules internalize through distinct pathways. Eur J Immunol. 1992 Jan;22(1):15–21. doi: 10.1002/eji.1830220104. [DOI] [PubMed] [Google Scholar]
- Bar-Sagi D., Feramisco J. R. Induction of membrane ruffling and fluid-phase pinocytosis in quiescent fibroblasts by ras proteins. Science. 1986 Sep 5;233(4768):1061–1068. doi: 10.1126/science.3090687. [DOI] [PubMed] [Google Scholar]
- Becich M. J., Mahklouf S., Baenziger J. U. Wheat germ agglutinin is selectively transported to multivesicular bodies. Eur J Cell Biol. 1991 Jun;55(1):83–93. [PubMed] [Google Scholar]
- Brown D., Orci L. Vasopressin stimulates formation of coated pits in rat kidney collecting ducts. Nature. 1983 Mar 17;302(5905):253–255. doi: 10.1038/302253a0. [DOI] [PubMed] [Google Scholar]
- Brown D., Weyer P., Orci L. Vasopressin stimulates endocytosis in kidney collecting duct principal cells. Eur J Cell Biol. 1988 Jun;46(2):336–341. [PubMed] [Google Scholar]
- Carlsson S. R., Roth J., Piller F., Fukuda M. Isolation and characterization of human lysosomal membrane glycoproteins, h-lamp-1 and h-lamp-2. Major sialoglycoproteins carrying polylactosaminoglycan. J Biol Chem. 1988 Dec 15;263(35):18911–18919. [PubMed] [Google Scholar]
- Geuze H. J., Slot J. W., van der Ley P. A., Scheffer R. C. Use of colloidal gold particles in double-labeling immunoelectron microscopy of ultrathin frozen tissue sections. J Cell Biol. 1981 Jun;89(3):653–665. doi: 10.1083/jcb.89.3.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldberg R. I., Smith R. M., Jarett L. Insulin and alpha 2-macroglobulin-methylamine undergo endocytosis by different mechanisms in rat adipocytes: I. Comparison of cell surface events. J Cell Physiol. 1987 Nov;133(2):203–212. doi: 10.1002/jcp.1041330202. [DOI] [PubMed] [Google Scholar]
- Goldstein J. L., Brown M. S., Anderson R. G., Russell D. W., Schneider W. J. Receptor-mediated endocytosis: concepts emerging from the LDL receptor system. Annu Rev Cell Biol. 1985;1:1–39. doi: 10.1146/annurev.cb.01.110185.000245. [DOI] [PubMed] [Google Scholar]
- Griffiths G., McDowall A., Back R., Dubochet J. On the preparation of cryosections for immunocytochemistry. J Ultrastruct Res. 1984 Oct;89(1):65–78. doi: 10.1016/s0022-5320(84)80024-6. [DOI] [PubMed] [Google Scholar]
- Haigler H. T., McKanna J. A., Cohen S. Rapid stimulation of pinocytosis in human carcinoma cells A-431 by epidermal growth factor. J Cell Biol. 1979 Oct;83(1):82–90. doi: 10.1083/jcb.83.1.82. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hansen S. H., Sandvig K., van Deurs B. Clathrin and HA2 adaptors: effects of potassium depletion, hypertonic medium, and cytosol acidification. J Cell Biol. 1993 Apr;121(1):61–72. doi: 10.1083/jcb.121.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hansen S. H., Sandvig K., van Deurs B. Internalization efficiency of the transferrin receptor. Exp Cell Res. 1992 Mar;199(1):19–28. doi: 10.1016/0014-4827(92)90457-j. [DOI] [PubMed] [Google Scholar]
- Hansen S. H., Sandvig K., van Deurs B. The preendosomal compartment comprises distinct coated and noncoated endocytic vesicle populations. J Cell Biol. 1991 May;113(4):731–741. doi: 10.1083/jcb.113.4.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hubbard A. L. Endocytosis. Curr Opin Cell Biol. 1989 Aug;1(4):675–683. doi: 10.1016/0955-0674(89)90033-1. [DOI] [PubMed] [Google Scholar]
- Huet C., Ash J. F., Singer S. J. The antibody-induced clustering and endocytosis of HLA antigens on cultured human fibroblasts. Cell. 1980 Sep;21(2):429–438. doi: 10.1016/0092-8674(80)90479-1. [DOI] [PubMed] [Google Scholar]
- Kartenbeck J., Schmelz M., Franke W. W., Geiger B. Endocytosis of junctional cadherins in bovine kidney epithelial (MDBK) cells cultured in low Ca2+ ion medium. J Cell Biol. 1991 May;113(4):881–892. doi: 10.1083/jcb.113.4.881. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kartenbeck J., Schmid E., Franke W. W., Geiger B. Different modes of internalization of proteins associated with adhaerens junctions and desmosomes: experimental separation of lateral contacts induces endocytosis of desmosomal plaque material. EMBO J. 1982;1(6):725–732. doi: 10.1002/j.1460-2075.1982.tb01237.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Keller G. A., Siegel M. W., Caras I. W. Endocytosis of glycophospholipid-anchored and transmembrane forms of CD4 by different endocytic pathways. EMBO J. 1992 Mar;11(3):863–874. doi: 10.1002/j.1460-2075.1992.tb05124.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Killisch I., Steinlein P., Römisch K., Hollinshead R., Beug H., Griffiths G. Characterization of early and late endocytic compartments of the transferrin cycle. Transferrin receptor antibody blocks erythroid differentiation by trapping the receptor in the early endosome. J Cell Sci. 1992 Sep;103(Pt 1):211–232. doi: 10.1242/jcs.103.1.211. [DOI] [PubMed] [Google Scholar]
- Larkin J. M., Brown M. S., Goldstein J. L., Anderson R. G. Depletion of intracellular potassium arrests coated pit formation and receptor-mediated endocytosis in fibroblasts. Cell. 1983 May;33(1):273–285. doi: 10.1016/0092-8674(83)90356-2. [DOI] [PubMed] [Google Scholar]
- Lencer W. I., Verkman A. S., Arnaout M. A., Ausiello D. A., Brown D. Endocytic vesicles from renal papilla which retrieve the vasopressin-sensitive water channel do not contain a functional H+ ATPase. J Cell Biol. 1990 Aug;111(2):379–389. doi: 10.1083/jcb.111.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lotti L. V., Di Lazzaro C., Zompetta C., Frati L., Torrisi M. R. Surface distribution and internalization of erbB-2 proteins. Exp Cell Res. 1992 Oct;202(2):274–280. doi: 10.1016/0014-4827(92)90075-j. [DOI] [PubMed] [Google Scholar]
- Madshus I. H., Sandvig K., Olsnes S., van Deurs B. Effect of reduced endocytosis induced by hypotonic shock and potassium depletion on the infection of Hep 2 cells by picornaviruses. J Cell Physiol. 1987 Apr;131(1):14–22. doi: 10.1002/jcp.1041310104. [DOI] [PubMed] [Google Scholar]
- Mellman I., Fuchs R., Helenius A. Acidification of the endocytic and exocytic pathways. Annu Rev Biochem. 1986;55:663–700. doi: 10.1146/annurev.bi.55.070186.003311. [DOI] [PubMed] [Google Scholar]
- Montesano R., Roth J., Robert A., Orci L. Non-coated membrane invaginations are involved in binding and internalization of cholera and tetanus toxins. Nature. 1982 Apr 15;296(5858):651–653. doi: 10.1038/296651a0. [DOI] [PubMed] [Google Scholar]
- Racoosin E. L., Swanson J. A. M-CSF-induced macropinocytosis increases solute endocytosis but not receptor-mediated endocytosis in mouse macrophages. J Cell Sci. 1992 Aug;102(Pt 4):867–880. doi: 10.1242/jcs.102.4.867. [DOI] [PubMed] [Google Scholar]
- Raposo G., Dunia I., Delavier-Klutchko C., Kaveri S., Strosberg A. D., Benedetti E. L. Internalization of beta-adrenergic receptor in A431 cells involves non-coated vesicles. Eur J Cell Biol. 1989 Dec;50(2):340–352. [PubMed] [Google Scholar]
- Ridley A. J., Paterson H. F., Johnston C. L., Diekmann D., Hall A. The small GTP-binding protein rac regulates growth factor-induced membrane ruffling. Cell. 1992 Aug 7;70(3):401–410. doi: 10.1016/0092-8674(92)90164-8. [DOI] [PubMed] [Google Scholar]
- Rodman J. S., Mercer R. W., Stahl P. D. Endocytosis and transcytosis. Curr Opin Cell Biol. 1990 Aug;2(4):664–672. doi: 10.1016/0955-0674(90)90108-q. [DOI] [PubMed] [Google Scholar]
- Sabolić I., Wuarin F., Shi L. B., Verkman A. S., Ausiello D. A., Gluck S., Brown D. Apical endosomes isolated from kidney collecting duct principal cells lack subunits of the proton pumping ATPase. J Cell Biol. 1992 Oct;119(1):111–122. doi: 10.1083/jcb.119.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sandvig K., van Deurs B. Selective modulation of the endocytic uptake of ricin and fluid phase markers without alteration in transferrin endocytosis. J Biol Chem. 1990 Apr 15;265(11):6382–6388. [PubMed] [Google Scholar]
- Severs N. J. Caveolae: static inpocketings of the plasma membrane, dynamic vesicles or plain artifact? J Cell Sci. 1988 Jul;90(Pt 3):341–348. doi: 10.1242/jcs.90.3.341. [DOI] [PubMed] [Google Scholar]
- Slot J. W., Geuze H. J. A new method of preparing gold probes for multiple-labeling cytochemistry. Eur J Cell Biol. 1985 Jul;38(1):87–93. [PubMed] [Google Scholar]
- Slot J. W., Geuze H. J., Gigengack S., Lienhard G. E., James D. E. Immuno-localization of the insulin regulatable glucose transporter in brown adipose tissue of the rat. J Cell Biol. 1991 Apr;113(1):123–135. doi: 10.1083/jcb.113.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spiegelberg H. L., Abel C. A., Fishkin B. G., Grey H. M. Localization of the carbohydrate within the variable region of light and heavy chains of human gamma g myeloma proteins. Biochemistry. 1970 Oct 13;9(21):4217–4223. doi: 10.1021/bi00823a025. [DOI] [PubMed] [Google Scholar]
- Stoorvogel W., Strous G. J., Geuze H. J., Oorschot V., Schwartz A. L. Late endosomes derive from early endosomes by maturation. Cell. 1991 May 3;65(3):417–427. doi: 10.1016/0092-8674(91)90459-c. [DOI] [PubMed] [Google Scholar]
- Strange K., Willingham M. C., Handler J. S., Harris H. W., Jr Apical membrane endocytosis via coated pits is stimulated by removal of antidiuretic hormone from isolated, perfused rabbit cortical collecting tubule. J Membr Biol. 1988 Jul;103(1):17–28. doi: 10.1007/BF01871929. [DOI] [PubMed] [Google Scholar]
- Tokuyasu K. T. Immunochemistry on ultrathin frozen sections. Histochem J. 1980 Jul;12(4):381–403. doi: 10.1007/BF01011956. [DOI] [PubMed] [Google Scholar]
- Tokuyasu K. T. Use of poly(vinylpyrrolidone) and poly(vinyl alcohol) for cryoultramicrotomy. Histochem J. 1989 Mar;21(3):163–171. doi: 10.1007/BF01007491. [DOI] [PubMed] [Google Scholar]
- Tran D., Carpentier J. L., Sawano F., Gorden P., Orci L. Ligands internalized through coated or noncoated invaginations follow a common intracellular pathway. Proc Natl Acad Sci U S A. 1987 Nov;84(22):7957–7961. doi: 10.1073/pnas.84.22.7957. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Verkman A. S., Lencer W. I., Brown D., Ausiello D. A. Endosomes from kidney collecting tubule cells contain the vasopressin-sensitive water channel. Nature. 1988 May 19;333(6170):268–269. doi: 10.1038/333268a0. [DOI] [PubMed] [Google Scholar]
- Watts C., Marsh M. Endocytosis: what goes in and how? J Cell Sci. 1992 Sep;103(Pt 1):1–8. doi: 10.1242/jcs.103.1.1a. [DOI] [PubMed] [Google Scholar]
- Weissman A. M., Klausner R. D., Rao K., Harford J. B. Exposure of K562 cells to anti-receptor monoclonal antibody OKT9 results in rapid redistribution and enhanced degradation of the transferrin receptor. J Cell Biol. 1986 Mar;102(3):951–958. doi: 10.1083/jcb.102.3.951. [DOI] [PMC free article] [PubMed] [Google Scholar]
- West M. A., Bretscher M. S., Watts C. Distinct endocytotic pathways in epidermal growth factor-stimulated human carcinoma A431 cells. J Cell Biol. 1989 Dec;109(6 Pt 1):2731–2739. doi: 10.1083/jcb.109.6.2731. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Deurs B., Holm P. K., Sandvig K., Hansen S. H. Are caveolae involved in clathrin-independent endocytosis? Trends Cell Biol. 1993 Aug;3(8):249–251. doi: 10.1016/0962-8924(93)90045-3. [DOI] [PubMed] [Google Scholar]
- van Deurs B., Petersen O. W., Olsnes S., Sandvig K. The ways of endocytosis. Int Rev Cytol. 1989;117:131–177. doi: 10.1016/s0074-7696(08)61336-4. [DOI] [PubMed] [Google Scholar]