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PMID: 9722606 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

An endocytosed TGN38 chimeric protein is delivered to the TGN after trafficking through the endocytic recycling compartment in CHO cells.

The Journal of cell biology ·Vol. 142 ·No. 4 ·1998-08-24 ·Pages 923-36

Ghosh RN, Mallet WG, Soe TT, McGraw TE, Maxfield FR

Abstract

To examine TGN38 trafficking from the cell surface to the TGN, CHO cells were stably transfected with a chimeric transmembrane protein, TacTGN38. We used fluorescent and 125I-labeled anti-Tac IgG and Fab fragments to follow TacTGN38's postendocytic trafficking. At steady-state, anti-Tac was mainly in the TGN, but shortly after endocytosis it was predominantly in early endosomes. 11% of cellular TacTGN38 is on the plasma membrane. Kinetic analysis of trafficking of antibodies bound to TacTGN38 showed that after short endocytic pulses, 80% of internalized anti-Tac returned to the cell surface (t1/2 = 9 min), and the remainder trafficked to the TGN. When longer filling pulses and chases were used to load anti-Tac into the TGN, it returned to the cell surface with a t1/2 of 46 min. Quantitative confocal microscopy analysis also showed that fluorescent anti-Tac fills the TGN with a 46-min t1/2. Using the measured rate constants in a simple kinetic model, we predict that 82% of TacTGN38 is in the TGN, and 7% is in endosomes. TacTGN38 leaves the TGN slowly, which accounts for its steady-state distribution despite the inefficient targeting from the cell surface to the TGN.

MeSH Terms
Animals Antibodies, Monoclonal/metabolism CHO Cells Cricetinae Endocytosis/physiology Endosomes/metabolism Glycoproteins Golgi Apparatus/physiology Kinetics Membrane Glycoproteins/metabolism Membrane Proteins/physiology Microscopy, Fluorescence Receptors, Interleukin-2/genetics,immunology Recombinant Fusion Proteins/metabolism Transfection/genetics Transferrin/metabolism
Chemicals
Antibodies, Monoclonal Glycoproteins Membrane Glycoproteins Membrane Proteins Receptors, Interleukin-2 Recombinant Fusion Proteins Transferrin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Ghosh R N
Department of Biochemistry, Cornell University Medical College, New York, New York 10021, USA.
Mallet W G
Soe T T
McGraw T E
Maxfield F R
References (52)
52 references, click to expand
  1. Rab11 regulates recycling through the pericentriolar recycling endosome.
    J Cell Biol. 1996 Nov;135(4):913-24 PMID: 8922376
  2. Evidence for nonvectorial, retrograde transferrin trafficking in the early endosomes of HEp2 cells.
    J Cell Biol. 1995 Feb;128(4):549-61 PMID: 7860630
  3. Kinetics of binding, endocytosis, and recycling of EGF receptor mutants.
    J Cell Biol. 1992 Apr;117(1):203-12 PMID: 1556153
  4. Structure and function of the mannose 6-phosphate/insulinlike growth factor II receptors.
    Annu Rev Biochem. 1992;61:307-30 PMID: 1323236
  5. The cytoplasmic domain mediates localization of furin to the trans-Golgi network en route to the endosomal/lysosomal system.
    J Cell Biol. 1994 Sep;126(5):1157-72 PMID: 7914893
  6. Endocytosis and lysosomal targeting of epidermal growth factor receptors are mediated by distinct sequences independent of the tyrosine kinase domain.
    J Biol Chem. 1995 Mar 3;270(9):4325-33 PMID: 7876194
  7. A His-Leu-Leu sequence near the carboxyl terminus of the cytoplasmic domain of the cation-dependent mannose 6-phosphate receptor is necessary for the lysosomal enzyme sorting function.
    J Biol Chem. 1992 Aug 25;267(24):17110-5 PMID: 1324923
  8. The TGN38 glycoprotein contains two non-overlapping signals that mediate localization to the trans-Golgi network.
    J Cell Biol. 1994 Apr;125(2):253-68 PMID: 8163544
  9. Transport of surface mannose 6-phosphate receptor to the Golgi complex in cultured human cells.
    J Biol Chem. 1989 May 5;264(13):7675-80 PMID: 2540200
  10. Functional expression of the human transferrin receptor cDNA in Chinese hamster ovary cells deficient in endogenous transferrin receptor.
    J Cell Biol. 1987 Jul;105(1):207-14 PMID: 3611186
  11. Point mutation at the ATP binding site of EGF receptor abolishes protein-tyrosine kinase activity and alters cellular routing.
    Cell. 1987 Oct 23;51(2):199-209 PMID: 3499230
  12. Quantification of low density lipoprotein and transferrin endocytic sorting HEp2 cells using confocal microscopy.
    J Cell Sci. 1994 Aug;107 ( Pt 8):2177-89 PMID: 7983176
  13. Low density lipoprotein receptor and cation-independent mannose 6-phosphate receptor are transported from the cell surface to the Golgi apparatus at equal rates in PC12 cells.
    J Cell Biol. 1992 Apr;117(1):47-55 PMID: 1313438
  14. Iterative fractionation of recycling receptors from lysosomally destined ligands in an early sorting endosome.
    J Cell Biol. 1989 Dec;109(6 Pt 2):3303-14 PMID: 2600137
  15. Cholesterol-dependent retention of GPI-anchored proteins in endosomes.
    EMBO J. 1998 Aug 17;17(16):4626-38 PMID: 9707422
  16. Localization of TGN38 to the trans-Golgi network: involvement of a cytoplasmic tyrosine-containing sequence.
    J Cell Biol. 1993 Mar;120(5):1123-35 PMID: 8436587
  17. Exocytosis of pinocytosed fluid in cultured cells: kinetic evidence for rapid turnover and compartmentation.
    J Cell Biol. 1981 Dec;91(3 Pt 1):716-27 PMID: 7328118
  18. Sorting of membrane components from endosomes and subsequent recycling to the cell surface occurs by a bulk flow process.
    J Cell Biol. 1993 Jun;121(6):1257-69 PMID: 8509447
  19. Endocytosis.
    Physiol Rev. 1997 Jul;77(3):759-803 PMID: 9234965
  20. The trans-Golgi network can be dissected structurally and functionally from the cisternae of the Golgi complex by brefeldin A.
    Eur J Cell Biol. 1992 Oct;59(1):92-105 PMID: 1468449
  21. The SXYQRL sequence in the cytoplasmic domain of TGN38 plays a major role in trans-Golgi network localization.
    J Biol Chem. 1993 Oct 25;268(30):22853-62 PMID: 8226795
  22. Intracellular movement of cell surface receptors after endocytosis: resialylation of asialo-transferrin receptor in human erythroleukemia cells.
    J Cell Biol. 1985 Mar;100(3):826-34 PMID: 2982885
  23. The End2 mutation in CHO cells slows the exit of transferrin receptors from the recycling compartment but bulk membrane recycling is unaffected.
    J Cell Biol. 1993 Sep;122(6):1231-41 PMID: 8376460
  24. Brefeldin A causes a microtubule-mediated fusion of the trans-Golgi network and early endosomes.
    Cell. 1991 Nov 1;67(3):591-600 PMID: 1657400
  25. TGN38 is maintained in the trans-Golgi network by a tyrosine-containing motif in the cytoplasmic domain.
    EMBO J. 1993 May;12(5):2219-28 PMID: 8491209
  26. The endocytotic rate constant. A cellular parameter for quantitating receptor-mediated endocytosis.
    J Biol Chem. 1982 Apr 25;257(8):4222-9 PMID: 6279628
  27. Identification, sequencing and expression of an integral membrane protein of the trans-Golgi network (TGN38).
    Biochem J. 1990 Aug 15;270(1):97-102 PMID: 2204342
  28. Membrane transport in the endocytic pathway.
    Curr Opin Cell Biol. 1995 Aug;7(4):552-63 PMID: 7495576
  29. Multiple pathways of exocytosis, endocytosis, and membrane recycling: validation of a Golgi route.
    Fed Proc. 1983 May 15;42(8):2407-13 PMID: 6404654
  30. Molecular trapping of a fluorescent ceramide analogue at the Golgi apparatus of fixed cells: interaction with endogenous lipids provides a trans-Golgi marker for both light and electron microscopy.
    J Cell Biol. 1989 Nov;109(5):2067-79 PMID: 2478562
  31. Delivery of ligands from sorting endosomes to late endosomes occurs by maturation of sorting endosomes.
    J Cell Biol. 1992 Apr;117(2):301-10 PMID: 1560027
  32. Oligomerized transferrin receptors are selectively retained by a lumenal sorting signal in a long-lived endocytic recycling compartment.
    J Cell Biol. 1995 Jun;129(6):1509-22 PMID: 7790351
  33. TGN38 and its orthologues: roles in post-TGN vesicle formation and maintenance of TGN morphology.
    Biochim Biophys Acta. 1997 Mar 1;1355(3):209-17 PMID: 9060992
  34. Ligand-mediated internalization, recycling, and downregulation of the epidermal growth factor receptor in vivo.
    J Cell Biol. 1989 Dec;109(6 Pt 1):2741-9 PMID: 2592403
  35. Two syntaxin homologues in the TGN/endosomal system of yeast.
    EMBO J. 1998 Jan 2;17(1):113-26 PMID: 9427746
  36. Rab11, a small GTPase associated with both constitutive and regulated secretory pathways in PC12 cells.
    FEBS Lett. 1993 Nov 15;334(2):175-82 PMID: 8224244
  37. Segregation of transferrin to a mildly acidic (pH 6.5) para-Golgi compartment in the recycling pathway.
    Cell. 1984 Jul;37(3):789-800 PMID: 6204769
  38. Intracellular trafficking and activation of the furin proprotein convertase: localization to the TGN and recycling from the cell surface.
    EMBO J. 1994 Jan 1;13(1):18-33 PMID: 7508380
  39. Human transferrin receptor internalization is partially dependent upon an aromatic amino acid on the cytoplasmic domain.
    Cell Regul. 1990 Mar;1(4):369-77 PMID: 2100204
  40. Intracellular fusion of sequentially formed endocytic compartments.
    J Cell Biol. 1988 Apr;106(4):1083-91 PMID: 3360848
  41. Fusion accessibility of endocytic compartments along the recycling and lysosomal endocytic pathways in intact cells.
    J Cell Biol. 1989 Nov;109(5):2097-104 PMID: 2681226
  42. An acidic sequence within the cytoplasmic domain of furin functions as a determinant of trans-Golgi network localization and internalization from the cell surface.
    EMBO J. 1995 Oct 16;14(20):4961-75 PMID: 7588625
  43. TGN38/41: a molecule on the move.
    Trends Cell Biol. 1993 Aug;3(8):252-5 PMID: 14731742
  44. An electron microscopic study of TGN38/41 dynamics.
    J Cell Sci Suppl. 1993;17:41-7 PMID: 8144704
  45. TGN38/41 recycles between the cell surface and the TGN: brefeldin A affects its rate of return to the TGN.
    Mol Biol Cell. 1993 Jan;4(1):93-105 PMID: 8443412
  46. Transferrin receptor containing the SDYQRL motif of TGN38 causes a reorganization of the recycling compartment but is not targeted to the TGN.
    J Cell Biol. 1996 Dec;135(6 Pt 2):1749-62 PMID: 8991088
  47. The cytoplasmic tail of the mannose 6-phosphate/insulin-like growth factor-II receptor has two signals for lysosomal enzyme sorting in the Golgi.
    J Cell Biol. 1992 Oct;119(2):249-57 PMID: 1400571
  48. Functional expression of furin demonstrating its intracellular localization and endoprotease activity for processing of proalbumin and complement pro-C3.
    J Biol Chem. 1991 Sep 5;266(25):16954-9 PMID: 1885622
  49. Insulin stimulation of GLUT-4 translocation: a model for regulated recycling.
    Trends Cell Biol. 1994 Apr;4(4):120-6 PMID: 14731734
  50. Brefeldin A's effects on endosomes, lysosomes, and the TGN suggest a general mechanism for regulating organelle structure and membrane traffic.
    Cell. 1991 Nov 1;67(3):601-16 PMID: 1682055
  51. Intracellular trafficking of furin is modulated by the phosphorylation state of a casein kinase II site in its cytoplasmic tail.
    EMBO J. 1995 Dec 1;14(23):5869-83 PMID: 8846780
  52. TGN38 recycles basolaterally in polarized Madin-Darby canine kidney cells.
    Mol Biol Cell. 1994 Oct;5(10):1093-103 PMID: 7865877
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.142.4.923
Published
1998-08-24
Pages
923-36
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2132871
Subset
IM
Grants
NIDDK NIH HHS · DK27083 · United States
External Links
PubMed source DOI: 10.1083/jcb.142.4.923 PMC Full Text View journal details
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