Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Jul 1;103(1):277–286. doi: 10.1083/jcb.103.1.277

Transferrin receptors recycle to cis and middle as well as trans Golgi cisternae in Ig-secreting myeloma cells

PMCID: PMC2113784  PMID: 3013900

Abstract

The recycling itinerary of plasma membrane transferrin receptors (TFR) was charted in IgG-secreting mouse myeloma cells (RPC 5.4) by tagging surface receptors with either bound anti-transferrin receptor antibodies (anti-TFR) or Fab fragments thereof and determining the intracellular destinations of the tagged receptors by immunocytochemistry. By immunofluorescence, TFR tagged with either probe were seen to be rapidly internalized and translocated from the cell surface to the juxtanuclear (Golgi) region. When localized by immunoperoxidase procedures at the electron microscopic level, the anti- TFR-labeled receptors were detected in all cisternae (cis, middle, and trans) of the Golgi stacks as well as in endosomes and trans Golgi reticular elements. There was no difference in the routing of TFR tagged with monovalent Fab and those tagged with divalent IgG. Tagged receptors were detected in Golgi stacks of approximately 50% of the cells analyzed. The position of the labeled cisternae within a given stack was found to be quite variable with cis and middle cisternae more often labeled at 5 min and trans cisternae at 30 min of antibody uptake. The finding that recycling plasmalemmal TFR can visit all or most Golgi subcompartments raises the likely possibility that any Golgi- associated posttranslational modification can occur during recycling as well as during the initial biosynthesis of plasmalemma receptors and other membrane proteins.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson R. G., Pathak R. K. Vesicles and cisternae in the trans Golgi apparatus of human fibroblasts are acidic compartments. Cell. 1985 Mar;40(3):635–643. doi: 10.1016/0092-8674(85)90212-0. [DOI] [PubMed] [Google Scholar]
  2. Brown W. J., Constantinescu E., Farquhar M. G. Redistribution of mannose-6-phosphate receptors induced by tunicamycin and chloroquine. J Cell Biol. 1984 Jul;99(1 Pt 1):320–326. doi: 10.1083/jcb.99.1.320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dautry-Varsat A., Ciechanover A., Lodish H. F. pH and the recycling of transferrin during receptor-mediated endocytosis. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2258–2262. doi: 10.1073/pnas.80.8.2258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dautry-Varsat A., Lodish H. F. How receptors bring proteins and particles into cells. Sci Am. 1984 May;250(5):52–58. doi: 10.1038/scientificamerican0584-52. [DOI] [PubMed] [Google Scholar]
  5. Dunphy W. G., Rothman J. E. Compartmentation of asparagine-linked oligosaccharide processing in the Golgi apparatus. J Cell Biol. 1983 Jul;97(1):270–275. doi: 10.1083/jcb.97.1.270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Farquhar M. G. Membrane recycling in secretory cells: implications for traffic of products and specialized membranes within the Golgi complex. Methods Cell Biol. 1981;23:399–427. doi: 10.1016/s0091-679x(08)61511-3. [DOI] [PubMed] [Google Scholar]
  7. Farquhar M. G. Membrane recycling in secretory cells: pathway to the Golgi complex. Ciba Found Symp. 1982;(92):157–183. doi: 10.1002/9780470720745.ch9. [DOI] [PubMed] [Google Scholar]
  8. Farquhar M. G. Multiple pathways of exocytosis, endocytosis, and membrane recycling: validation of a Golgi route. Fed Proc. 1983 May 15;42(8):2407–2413. [PubMed] [Google Scholar]
  9. Farquhar M. G. Progress in unraveling pathways of Golgi traffic. Annu Rev Cell Biol. 1985;1:447–488. doi: 10.1146/annurev.cb.01.110185.002311. [DOI] [PubMed] [Google Scholar]
  10. Farquhar M. G. Recovery of surface membrane in anterior pituitary cells. Variations in traffic detected with anionic and cationic ferritin. J Cell Biol. 1978 Jun;77(3):R35–R42. doi: 10.1083/jcb.77.3.r35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldberg D. E., Kornfeld S. Evidence for extensive subcellular organization of asparagine-linked oligosaccharide processing and lysosomal enzyme phosphorylation. J Biol Chem. 1983 Mar 10;258(5):3159–3165. [PubMed] [Google Scholar]
  12. Harding C., Heuser J., Stahl P. Receptor-mediated endocytosis of transferrin and recycling of the transferrin receptor in rat reticulocytes. J Cell Biol. 1983 Aug;97(2):329–339. doi: 10.1083/jcb.97.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hausen P., Dreyer C. The use of polyacrylamide as an embedding medium for immunohistochemical studies of embryonic tissues. Stain Technol. 1981 Sep;56(5):287–293. doi: 10.3109/10520298109067329. [DOI] [PubMed] [Google Scholar]
  14. Herzog V., Farquhar M. G. Luminal membrane retrieved after exocytosis reaches most golgi cisternae in secretory cells. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5073–5077. doi: 10.1073/pnas.74.11.5073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Herzog V., Miller F. Membrane retrieval in epithelial cells of isolated thyroid follicles. Eur J Cell Biol. 1979 Aug;19(3):203–215. [PubMed] [Google Scholar]
  16. Hopkins C. R. Intracellular routing of transferrin and transferrin receptors in epidermoid carcinoma A431 cells. Cell. 1983 Nov;35(1):321–330. doi: 10.1016/0092-8674(83)90235-0. [DOI] [PubMed] [Google Scholar]
  17. Hopkins C. R., Trowbridge I. S. Internalization and processing of transferrin and the transferrin receptor in human carcinoma A431 cells. J Cell Biol. 1983 Aug;97(2):508–521. doi: 10.1083/jcb.97.2.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Keller G. A., Tokuyasu K. T., Dutton A. H., Singer S. J. An improved procedure for immunoelectron microscopy: ultrathin plastic embedding of immunolabeled ultrathin frozen sections. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5744–5747. doi: 10.1073/pnas.81.18.5744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Klausner R. D., Ashwell G., van Renswoude J., Harford J. B., Bridges K. R. Binding of apotransferrin to K562 cells: explanation of the transferrin cycle. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2263–2266. doi: 10.1073/pnas.80.8.2263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kornfeld R., Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–664. doi: 10.1146/annurev.bi.54.070185.003215. [DOI] [PubMed] [Google Scholar]
  21. Lesley J. F., Schulte R. J. Inhibition of cell growth by monoclonal anti-transferrin receptor antibodies. Mol Cell Biol. 1985 Aug;5(8):1814–1821. doi: 10.1128/mcb.5.8.1814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lesley J. F., Schulte R. J. Selection of cell lines resistant to anti-transferrin receptor antibody: evidence for a mutation in transferrin receptor. Mol Cell Biol. 1984 Sep;4(9):1675–1681. doi: 10.1128/mcb.4.9.1675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mazia D., Schatten G., Sale W. Adhesion of cells to surfaces coated with polylysine. Applications to electron microscopy. J Cell Biol. 1975 Jul;66(1):198–200. doi: 10.1083/jcb.66.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
  25. Mellman I., Plutner H. Internalization and degradation of macrophage Fc receptors bound to polyvalent immune complexes. J Cell Biol. 1984 Apr;98(4):1170–1177. doi: 10.1083/jcb.98.4.1170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ottosen P. D., Courtoy P. J., Farquhar M. G. Pathways followed by membrane recovered from the surface of plasma cells and myeloma cells. J Exp Med. 1980 Jul 1;152(1):1–19. doi: 10.1084/jem.152.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Platt J. L., Michael A. F. Retardation of fading and enhancement of intensity of immunofluorescence by p-phenylenediamine. J Histochem Cytochem. 1983 Jun;31(6):840–842. doi: 10.1177/31.6.6341464. [DOI] [PubMed] [Google Scholar]
  28. Pohlmann R., Waheed A., Hasilik A., von Figura K. Synthesis of phosphorylated recognition marker in lysosomal enzymes is located in the cis part of Golgi apparatus. J Biol Chem. 1982 May 25;257(10):5323–5325. [PubMed] [Google Scholar]
  29. Regoeczi E., Chindemi P. A., Debanne M. T., Charlwood P. A. Partial resialylation of human asialotransferrin type 3 in the rat. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2226–2230. doi: 10.1073/pnas.79.7.2226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Regoeczi E., Chindemi P. A., Debanne M. T. Partial resialylation of human asialotransferrin types 1 and 2 in the rat. Can J Biochem Cell Biol. 1984 Sep;62(9):853–858. doi: 10.1139/o84-109. [DOI] [PubMed] [Google Scholar]
  31. Roth J., Taatjes D. J., Lucocq J. M., Weinstein J., Paulson J. C. Demonstration of an extensive trans-tubular network continuous with the Golgi apparatus stack that may function in glycosylation. Cell. 1985 Nov;43(1):287–295. doi: 10.1016/0092-8674(85)90034-0. [DOI] [PubMed] [Google Scholar]
  32. Segal D. M., Hurwitz E. Binding of affinity cross-linked oligomers of IgG to cells bearing Fc receptors. J Immunol. 1977 Apr;118(4):1338–1337. [PubMed] [Google Scholar]
  33. Snider M. D., Rogers O. C. Intracellular movement of cell surface receptors after endocytosis: resialylation of asialo-transferrin receptor in human erythroleukemia cells. J Cell Biol. 1985 Mar;100(3):826–834. doi: 10.1083/jcb.100.3.826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Trowbridge I. S., Newman R. A., Domingo D. L., Sauvage C. Transferrin receptors: structure and function. Biochem Pharmacol. 1984 Mar 15;33(6):925–932. doi: 10.1016/0006-2952(84)90447-7. [DOI] [PubMed] [Google Scholar]
  35. Tycko B., Maxfield F. R. Rapid acidification of endocytic vesicles containing alpha 2-macroglobulin. Cell. 1982 Mar;28(3):643–651. doi: 10.1016/0092-8674(82)90219-7. [DOI] [PubMed] [Google Scholar]
  36. Willingham M. C., Pastan I. Ultrastructural immunocytochemical localization of the transferrin receptor using a monoclonal antibody in human KB cells. J Histochem Cytochem. 1985 Jan;33(1):59–64. doi: 10.1177/33.1.2856926. [DOI] [PubMed] [Google Scholar]
  37. Yamashiro D. J., Tycko B., Fluss S. R., Maxfield F. R. Segregation of transferrin to a mildly acidic (pH 6.5) para-Golgi compartment in the recycling pathway. Cell. 1984 Jul;37(3):789–800. doi: 10.1016/0092-8674(84)90414-8. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES