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PMID: 8741847 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The paramyxovirus simian virus 5 hemagglutinin-neuraminidase glycoprotein, but not the fusion glycoprotein, is internalized via coated pits and enters the endocytic pathway.

Molecular biology of the cell ·Vol. 7 ·No. 1 ·1996-01-00 ·Pages 155-72

Leser GP, Ector KJ, Lamb RA

Abstract

The hemagglutinin-neuraminidase (HN) and fusion (F) glycoproteins of the paramyxovirus simian virus 5 (SV5) are expressed on the surface of virus-infected cells. Although the F protein was found to be expressed stably, the HN protein was internalized from the plasma membrane. HN protein lacks known internalization signals in its cytoplasmic domain that are common to many integral membrane proteins that are internalized via clathrin-coated pits. Thus, the cellular pathway of HN protein internalization was examined. Biochemical analysis indicated that HN was lost from the cell surface with a t1/2 of approximately 45-50 min and turned over with a t1/2 of approximately 2 h. Immunofluorescent analysis showed internalized SV5 HN in vesicle-like structures in a juxtanuclear pattern coincident with the localization of ovalbumin. In contrast the SV5 F glycoprotein and the HN glycoprotein of the highly related parainfluenza virus 3 (hPIV-3) were found only on the cell surface. Immunogold staining of HN on the surface of SV5-infected CV-1 cells and examination using electron microscopy, showed heavy surface labeling that gradually decreased with time. Concomitantly, gold particles were detected in the endosomal system and with increasing time, gold-labeled structures having the morphology of lysosomes were observed. On the plasma membrane approximately 5% of the gold-labeled HN was found in coated pits. The inhibition of the pinching-off of coated pits from the plasma membrane by cytosol acidification significantly reduced HN internalization. Internalized HN was co-localized with gold-conjugated transferrin, a marker for the early endosomal compartments, and with gold-conjugated bovine serum albumin, a marker for late endosomal compartments. Taken together, these data strongly suggest that the HN glycoprotein is internalized via clathrin-coated pits and delivered to the endocytic pathway.

MeSH Terms
Cell Line Coated Pits, Cell-Membrane/virology Endocytosis Fluorescent Antibody Technique, Indirect HN Protein/metabolism Immunohistochemistry Microscopy, Electron Respirovirus/chemistry Viral Fusion Proteins/metabolism
Chemicals
HN Protein Viral Fusion Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Leser G P
Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, Illinois 60208-3500, USA.
Ector K J
Lamb R A
References (85)
85 references, click to expand
  1. Intracellular maturation and transport of the SV5 type II glycoprotein hemagglutinin-neuraminidase: specific and transient association with GRP78-BiP in the endoplasmic reticulum and extensive internalization from the cell surface.
    J Cell Biol. 1989 Dec;109(6 Pt 2):3273-89 PMID: 2557352
  2. Identification of a novel sequence mediating regulated endocytosis of the G protein-coupled alpha-pheromone receptor in yeast.
    Mol Biol Cell. 1993 May;4(5):511-21 PMID: 8392878
  3. Non-coated membrane invaginations are involved in binding and internalization of cholera and tetanus toxins.
    Nature. 1982 Apr 15;296(5858):651-3 PMID: 7070509
  4. Cell surface expression of glycosylated, nonglycosylated, and truncated forms of a cytoplasmic protein pyruvate kinase.
    J Cell Biol. 1988 Sep;107(3):865-76 PMID: 3047152
  5. A single amino acid change in the cytoplasmic domain allows the influenza virus hemagglutinin to be endocytosed through coated pits.
    Cell. 1988 Jun 3;53(5):743-52 PMID: 2897244
  6. Coated endosomal vesicles: sorting and recycling compartment for transferrin in BHK cells.
    Eur J Cell Biol. 1991 Dec;56(2):210-22 PMID: 1802708
  7. Signal-dependent membrane protein trafficking in the endocytic pathway.
    Annu Rev Cell Biol. 1993;9:129-61 PMID: 8280459
  8. Endocytosis.
    Curr Opin Cell Biol. 1989 Aug;1(4):675-83 PMID: 2516741
  9. Characteristics of the tyrosine recognition signal for internalization of transmembrane surface glycoproteins.
    J Cell Biol. 1990 Oct;111(4):1393-407 PMID: 2120240
  10. Kinase activity controls the sorting of the epidermal growth factor receptor within the multivesicular body.
    Cell. 1990 May 18;61(4):623-34 PMID: 2344614
  11. Intracellular transport of transferrin- and asialoorosomucoid-colloidal gold conjugates to lysosomes after receptor-mediated endocytosis.
    J Histochem Cytochem. 1985 Nov;33(11):1134-44 PMID: 2997327
  12. A glycolipid-anchored prion protein is endocytosed via clathrin-coated pits.
    J Cell Biol. 1994 Jun;125(6):1239-50 PMID: 7911471
  13. Mutagenesis of the human transferrin receptor: two cytoplasmic phenylalanines are required for efficient internalization and a second-site mutation is capable of reverting an internalization-defective phenotype.
    J Cell Biol. 1991 Mar;112(5):853-61 PMID: 1900298
  14. Endocytosis of glycophospholipid-anchored and transmembrane forms of CD4 by different endocytic pathways.
    EMBO J. 1992 Mar;11(3):863-74 PMID: 1532143
  15. Specific binding sites for albumin restricted to plasmalemmal vesicles of continuous capillary endothelium: receptor-mediated transcytosis.
    J Cell Biol. 1986 Apr;102(4):1304-11 PMID: 3007533
  16. Folding and assembly of viral membrane proteins.
    Virology. 1993 Apr;193(2):545-62 PMID: 8460475
  17. Basolateral sorting in MDCK cells requires a distinct cytoplasmic domain determinant.
    Cell. 1991 Sep 6;66(5):907-20 PMID: 1909606
  18. Isolation and characterization of monoclonal antibodies to simian virus 5 and their use in revealing antigenic differences between human, canine and simian isolates.
    J Gen Virol. 1987 Nov;68 ( Pt 11):2769-80 PMID: 2445904
  19. Endocytosis of the transferrin receptor requires the cytoplasmic domain but not its phosphorylation site.
    Cell. 1987 May 8;49(3):423-31 PMID: 3568132
  20. Receptors compete for adaptors found in plasma membrane coated pits.
    EMBO J. 1988 Nov;7(11):3331-6 PMID: 2905261
  21. M-CSF-induced macropinocytosis increases solute endocytosis but not receptor-mediated endocytosis in mouse macrophages.
    J Cell Sci. 1992 Aug;102 ( Pt 4):867-80 PMID: 1429898
  22. Vesicular stomatitis virus glycoprotein is sorted and concentrated during export from the endoplasmic reticulum.
    Cell. 1994 Mar 11;76(5):841-52 PMID: 8124720
  23. Endocytosis: what goes in and how?
    J Cell Sci. 1992 Sep;103 ( Pt 1):1-8 PMID: 1429899
  24. Hemagglutinin-neuraminidase protein of the paramyxovirus simian virus 5: nucleotide sequence of the mRNA predicts an N-terminal membrane anchor.
    J Virol. 1985 Apr;54(1):1-6 PMID: 3973974
  25. The glycophospholipid-linked folate receptor internalizes folate without entering the clathrin-coated pit endocytic pathway.
    J Cell Biol. 1990 Mar;110(3):637-49 PMID: 1968465
  26. Deletions in the cytoplasmic domain of the polymeric immunoglobulin receptor differentially affect endocytotic rate and postendocytotic traffic.
    J Biol Chem. 1990 Aug 15;265(23):13750-7 PMID: 2380185
  27. Heterologous transmembrane and cytoplasmic domains direct functional chimeric influenza virus hemagglutinins into the endocytic pathway.
    J Cell Biol. 1986 Apr;102(4):1271-83 PMID: 3007532
  28. Glycolipid-anchored proteins in neuroblastoma cells form detergent-resistant complexes without caveolin.
    J Cell Biol. 1995 May;129(3):619-27 PMID: 7537273
  29. Clathrin facilitates the internalization of seven transmembrane segment receptors for mating pheromones in yeast.
    J Cell Biol. 1993 Dec;123(6 Pt 2):1707-16 PMID: 8276891
  30. Consumption of EGF by A431 cells: evidence for receptor recycling.
    J Cell Biol. 1993 Jan;120(1):85-93 PMID: 8416997
  31. Continuous internalization of tumor necrosis factor receptors in a human myosarcoma cell line.
    J Biol Chem. 1988 Jul 25;263(21):10262-6 PMID: 2839482
  32. Transferrin receptors and cation-independent mannose-6-phosphate receptors deliver their ligands to two distinct subpopulations of multivesicular endosomes.
    Eur J Cell Biol. 1989 Oct;50(1):132-43 PMID: 2558886
  33. Are caveolae involved in clathrin-independent endocytosis?
    Trends Cell Biol. 1993 Aug;3(8):249-51 PMID: 14731741
  34. Transferrin receptor internalization sequence YXRF implicates a tight turn as the structural recognition motif for endocytosis.
    Cell. 1990 Nov 30;63(5):1061-72 PMID: 2257624
  35. A new method of preparing gold probes for multiple-labeling cytochemistry.
    Eur J Cell Biol. 1985 Jul;38(1):87-93 PMID: 4029177
  36. Distinct endocytotic pathways in epidermal growth factor-stimulated human carcinoma A431 cells.
    J Cell Biol. 1989 Dec;109(6 Pt 1):2731-9 PMID: 2556406
  37. Fc receptor isoforms exhibit distinct abilities for coated pit localization as a result of cytoplasmic domain heterogeneity.
    Cell. 1989 Jul 28;58(2):317-27 PMID: 2568890
  38. Induction of caveolin during adipogenesis and association of GLUT4 with caveolin-rich vesicles.
    J Cell Biol. 1994 Dec;127(5):1233-43 PMID: 7962086
  39. Antigenic and functional organization of human parainfluenza virus type 3 fusion glycoprotein.
    J Virol. 1989 Jan;63(1):375-82 PMID: 2462062
  40. Vectorial targeting of apical and basolateral plasma membrane proteins in a human adenocarcinoma epithelial cell line.
    Proc Natl Acad Sci U S A. 1989 Dec;86(23):9313-7 PMID: 2687880
  41. The internalization signal in the cytoplasmic tail of lysosomal acid phosphatase consists of the hexapeptide PGYRHV.
    EMBO J. 1992 Dec;11(12):4391-9 PMID: 1425575
  42. The J.D. mutation in familial hypercholesterolemia: amino acid substitution in cytoplasmic domain impedes internalization of LDL receptors.
    Cell. 1986 Apr 11;45(1):15-24 PMID: 3955657
  43. Annexin I is phosphorylated in the multivesicular body during the processing of the epidermal growth factor receptor.
    J Cell Biol. 1993 Jan;120(1):77-83 PMID: 8093248
  44. NPXY, a sequence often found in cytoplasmic tails, is required for coated pit-mediated internalization of the low density lipoprotein receptor.
    J Biol Chem. 1990 Feb 25;265(6):3116-23 PMID: 1968060
  45. The cytoplasmic domain of the diphtheria toxin receptor (HB-EGF precursor) is not required for receptor-mediated endocytosis.
    J Biol Chem. 1994 Oct 28;269(43):26635-41 PMID: 7929396
  46. Role of the human transferrin receptor cytoplasmic domain in endocytosis: localization of a specific signal sequence for internalization.
    J Cell Biol. 1990 Feb;110(2):283-94 PMID: 2298808
  47. The protein tyrosine kinase p56lck inhibits CD4 endocytosis by preventing entry of CD4 into coated pits.
    J Cell Biol. 1992 Apr;117(2):279-90 PMID: 1373141
  48. A quantitative model of traffic between plasma membrane and secondary lysosomes: evaluation of inflow, lateral diffusion, and degradation.
    J Cell Biol. 1988 Dec;107(6 Pt 1):2109-15 PMID: 2848849
  49. A point mutation in the cytoplasmic domain of the transferrin receptor inhibits endocytosis.
    Biochem J. 1990 Apr 1;267(1):31-5 PMID: 2327986
  50. The NPXY internalization signal of the LDL receptor adopts a reverse-turn conformation.
    Cell. 1991 Dec 20;67(6):1195-201 PMID: 1760844
  51. Endocytosis of the ASGP receptor H1 is reduced by mutation of tyrosine-5 but still occurs via coated pits.
    J Cell Biol. 1991 Aug;114(3):423-31 PMID: 1907285
  52. Cytosol- and clathrin-dependent stimulation of endocytosis in vitro by purified adaptors.
    J Cell Biol. 1992 Dec;119(5):1163-71 PMID: 1447294
  53. Multivesicular bodies in HEp-2 cells are maturing endosomes.
    Eur J Cell Biol. 1993 Aug;61(2):208-24 PMID: 8223712
  54. Calcium pump of the plasma membrane is localized in caveolae.
    J Cell Biol. 1993 Mar;120(5):1147-57 PMID: 8382206
  55. Cycling of the integral membrane glycoprotein, LEP100, between plasma membrane and lysosomes: kinetic and morphological analysis.
    Cell. 1987 Jun 5;49(5):669-77 PMID: 3107839
  56. Fusion protein of the paramyxovirus simian virus 5: nucleotide sequence of mRNA predicts a highly hydrophobic glycoprotein.
    Proc Natl Acad Sci U S A. 1984 Nov;81(21):6706-10 PMID: 6093114
  57. Mutations in the cytoplasmic domain of the 275 kd mannose 6-phosphate receptor differentially alter lysosomal enzyme sorting and endocytosis.
    Cell. 1989 Jun 2;57(5):787-96 PMID: 2541923
  58. 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
  59. Specificity of binding of clathrin adaptors to signals on the mannose-6-phosphate/insulin-like growth factor II receptor.
    EMBO J. 1989 Apr;8(4):1041-7 PMID: 2545438
  60. The basolateral targeting signal in the cytoplasmic domain of glycoprotein G from vesicular stomatitis virus resembles a variety of intracellular targeting motifs related by primary sequence but having diverse targeting activities.
    J Biol Chem. 1994 Jun 3;269(22):15732-9 PMID: 8195226
  61. Role of the coated endocytic vesicle in the uptake of receptor-bound low density lipoprotein in human fibroblasts.
    Cell. 1977 Mar;10(3):351-64 PMID: 191195
  62. Differential endocytosis of CD4 in lymphocytic and nonlymphocytic cells.
    J Exp Med. 1991 Mar 1;173(3):575-87 PMID: 1900077
  63. A quantitative analysis of the endocytic pathway in baby hamster kidney cells.
    J Cell Biol. 1989 Dec;109(6 Pt 1):2703-20 PMID: 2592402
  64. Vesicle fusion and formation at the surface of pinocytic vacuoles in macrophages.
    J Cell Biol. 1968 Sep;38(3):629-32 PMID: 5664229
  65. Internalization efficiency of the transferrin receptor.
    Exp Cell Res. 1992 Mar;199(1):19-28 PMID: 1735458
  66. A mutation that impairs the ability of lipoprotein receptors to localise in coated pits on the cell surface of human fibroblasts.
    Nature. 1977 Dec 22-29;270(5639):695-9 PMID: 201867
  67. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy.
    J Histochem Cytochem. 1974 Dec;22(12):1077-83 PMID: 4374474
  68. Vesicular stomatitis virus glycoprotein contains a dominant cytoplasmic basolateral sorting signal critically dependent upon a tyrosine.
    J Biol Chem. 1993 Feb 15;268(5):3313-20 PMID: 8381425
  69. Regulated internalization of caveolae.
    J Cell Biol. 1994 Dec;127(5):1199-215 PMID: 7962085
  70. Lack of detectable endocytosis of B lymphocyte MHC class II antigens using an antibody-independent technique.
    J Immunol. 1990 Feb 1;144(3):990-7 PMID: 2295824
  71. Circulating integrins: alpha 5 beta 1, alpha 6 beta 4 and Mac-1, but not alpha 3 beta 1, alpha 4 beta 1 or LFA-1.
    EMBO J. 1992 Feb;11(2):405-10 PMID: 1531629
  72. The low density lipoprotein receptor. Identification of amino acids in cytoplasmic domain required for rapid endocytosis.
    J Biol Chem. 1987 Mar 25;262(9):4075-82 PMID: 3104336
  73. A role for the cytoplasmic domain in transferrin receptor sorting and coated pit formation during endocytosis.
    Cell. 1988 Aug 12;54(4):485-9 PMID: 2900073
  74. Receptor-mediated endocytosis of interleukin 2 in a human tumor T cell line. Degradation of interleukin 2 and evidence for the absence of recycling of interleukin receptors.
    J Biol Chem. 1986 Nov 25;261(33):15450-4 PMID: 3096988
  75. Transferrin receptors promote the formation of clathrin lattices.
    Cell. 1991 May 17;65(4):621-32 PMID: 1903330
  76. Stage-specific assays for coated pit formation and coated vesicle budding in vitro.
    J Cell Biol. 1991 Sep;114(5):869-80 PMID: 1908470
  77. Internalization-defective LDL receptors produced by genes with nonsense and frameshift mutations that truncate the cytoplasmic domain.
    Cell. 1985 Jul;41(3):735-43 PMID: 3924410
  78. Acidification of the cytosol inhibits endocytosis from coated pits.
    J Cell Biol. 1987 Aug;105(2):679-89 PMID: 2887575
  79. Internalization and processing of transferrin and the transferrin receptor in human carcinoma A431 cells.
    J Cell Biol. 1983 Aug;97(2):508-21 PMID: 6309862
  80. Endocytosis and the recycling of plasma membrane.
    J Cell Biol. 1983 Jan;96(1):1-27 PMID: 6298247
  81. Characterization of the budding compartment of mouse hepatitis virus: evidence that transport from the RER to the Golgi complex requires only one vesicular transport step.
    J Cell Biol. 1994 Jan;124(1-2):55-70 PMID: 8294506
  82. Induction of membrane ruffling and fluid-phase pinocytosis in quiescent fibroblasts by ras proteins.
    Science. 1986 Sep 5;233(4768):1061-8 PMID: 3090687
  83. Transepithelial transport of a viral membrane glycoprotein implanted into the apical plasma membrane of Madin-Darby canine kidney cells. I. Morphological evidence.
    J Cell Biol. 1983 Sep;97(3):627-37 PMID: 6885914
  84. The rubella virus E2 and E1 spike glycoproteins are targeted to the Golgi complex.
    J Cell Biol. 1993 Apr;121(2):269-81 PMID: 8468347
  85. Dual pathways of internalization of the cholecystokinin receptor.
    J Cell Biol. 1995 Mar;128(6):1029-41 PMID: 7896869
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.7.1.155
Published
1996-01-00
Pages
155-72
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC278620
Subset
IM
Grants
NIAID NIH HHS · AI-23173 · United States
External Links
PubMed source DOI: 10.1091/mbc.7.1.155 PMC Full Text View journal details
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