Home LiteratureArticle Details
PMID: 8612570 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Reconstitution of transcytosis in SLO-permeabilized MDCK cells: existence of an NSF-dependent fusion mechanism with the apical surface of MDCK cells.

The EMBO journal ·Vol. 15 ·No. 7 ·1996-04-01 ·Pages 1471-81

Apodaca G, Cardone MH, Whiteheart SW, DasGupta BR, Mostov KE

Abstract

Recently, it was demonstrated that delivery from the trans-Golgi network (TGN) to the basolateral surface of Madin-Darby canine kidney (MDCK) cells required N-ethylmaleimide-sensitive factor (NSF)-alpha soluble NSF attachment protein (SNAP)-SNAP receptor (SNARE) complexes, while delivery from the TGN to the apical surface was independent of NSF-alpha SNAP-SNARE. To determine if all traffic to the apical surface of this cell line was NSF independent, we reconstituted the transcytosis of pre-internalized IgA to the apical surface and recycling to the basolateral surface. Transcytosis and the recycling of IgA required ATP and cytosol, and both were inhibited by treatment with N-ethylmaleimide. This inhibition was reversed by the addition of recombinant NSF. Botulinum neurotoxin serotype E, which is known to cleave the 25,000 Da synaptosomal associated protein, inhibited both transcytosis and recycling, although incompletely. We conclude that membrane traffic to a target membrane is not determined by utilizing a single molecular mechanism for fusion. Rather, a target membrane, e.g. the apical plasma membrane of MDCK cells, may use multiple molecular mechanisms to fuse with incoming vesicle.

MeSH Terms
Adenosine Triphosphate/metabolism Animals Bacterial Proteins Biological Transport, Active/drug effects Botulinum Toxins/pharmacology Carrier Proteins/metabolism Cell Line Cell Membrane Permeability Cytosol/metabolism Dogs Enzyme Inhibitors/pharmacology Ethers, Cyclic/pharmacology Golgi Apparatus/drug effects,metabolism Immunoglobulin A/metabolism In Vitro Techniques Membrane Fusion/physiology Membrane Proteins/metabolism Microtubules/drug effects Models, Biological N-Ethylmaleimide-Sensitive Proteins Nocodazole/pharmacology Okadaic Acid Phosphoprotein Phosphatases/antagonists & inhibitors Rats Rats, Sprague-Dawley SNARE Proteins Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins Streptolysins Vesicular Transport Proteins
Chemicals
Bacterial Proteins Carrier Proteins Enzyme Inhibitors Ethers, Cyclic Immunoglobulin A Membrane Proteins SNARE Proteins Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins Streptolysins Vesicular Transport Proteins streptolysin O Okadaic Acid Adenosine Triphosphate Phosphoprotein Phosphatases Botulinum Toxins N-Ethylmaleimide-Sensitive Proteins Nsf protein, rat Nocodazole
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Apodaca G
Department of Anatomy, University of California, San Francisco 94143-0452, USA.
Cardone M H
Whiteheart S W
DasGupta B R
Mostov K E
References (59)
59 references, click to expand
  1. Intracellular pools of transferrin receptors result from constitutive internalization of unoccupied receptors.
    Proc Natl Acad Sci U S A. 1986 Sep;83(17):6445-9 PMID: 2875458
  2. ATP and cytosol requirements for transferrin recycling in intact and disrupted MDCK cells.
    EMBO J. 1990 Nov;9(11):3477-87 PMID: 2209555
  3. The yeast SEC17 gene product is functionally equivalent to mammalian alpha-SNAP protein.
    J Biol Chem. 1992 Jun 15;267(17):12106-15 PMID: 1601878
  4. Expression and analysis of the polymeric immunoglobulin receptor in Madin-Darby canine kidney cells using retroviral vectors.
    Methods Cell Biol. 1989;32:329-37 PMID: 2532705
  5. Proteolysis of SNAP-25 by types E and A botulinal neurotoxins.
    J Biol Chem. 1994 Jan 21;269(3):1617-20 PMID: 8294407
  6. Annexin XIIIb: a novel epithelial specific annexin is implicated in vesicular traffic to the apical plasma membrane.
    J Cell Biol. 1995 Mar;128(6):1043-53 PMID: 7896870
  7. SNAP receptors implicated in vesicle targeting and fusion.
    Nature. 1993 Mar 25;362(6418):318-24 PMID: 8455717
  8. SNAP-25 is expressed in islets of Langerhans and is involved in insulin release.
    J Cell Biol. 1995 Mar;128(6):1019-28 PMID: 7896868
  9. Separation, purification, partial characterization and comparison of the heavy and light chains of botulinum neurotoxin types A, B, and E.
    J Biol Chem. 1985 Sep 5;260(19):10461-6 PMID: 4030755
  10. Homologs of the synaptobrevin/VAMP family of synaptic vesicle proteins function on the late secretory pathway in S. cerevisiae.
    Cell. 1993 Sep 10;74(5):855-61 PMID: 8374953
  11. The polymeric immunoglobulin receptor. A model protein to study transcytosis.
    J Clin Invest. 1991 Jun;87(6):1877-82 PMID: 2040683
  12. Basolateral protein transport in streptolysin O-permeabilized MDCK cells.
    J Cell Biol. 1994 Jun;125(5):1025-35 PMID: 8195286
  13. Purification of an N-ethylmaleimide-sensitive protein catalyzing vesicular transport.
    Proc Natl Acad Sci U S A. 1988 Nov;85(21):7852-6 PMID: 3186695
  14. Gs alpha stimulates transcytosis and apical secretion in MDCK cells through cAMP and protein kinase A.
    J Cell Biol. 1994 Aug;126(3):677-87 PMID: 8045932
  15. SNAPs and NSF: general members of the fusion apparatus.
    Trends Cell Biol. 1995 Feb;5(2):64-8 PMID: 14731414
  16. The formation of Golgi stacks from vesiculated Golgi membranes requires two distinct fusion events.
    Cell. 1995 Sep 22;82(6):895-904 PMID: 7553850
  17. Differential microtubule requirements for transcytosis in MDCK cells.
    EMBO J. 1990 Nov;9(11):3515-25 PMID: 2170116
  18. Sec9 is a SNAP-25-like component of a yeast SNARE complex that may be the effector of Sec4 function in exocytosis.
    Cell. 1994 Oct 21;79(2):245-58 PMID: 7954793
  19. 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
  20. Different requirements for NSF, SNAP, and Rab proteins in apical and basolateral transport in MDCK cells.
    Cell. 1995 May 19;81(4):571-80 PMID: 7758111
  21. The karyogamy gene KAR2 and novel proteins are required for ER-membrane fusion.
    Cell. 1994 Jul 15;78(1):87-98 PMID: 8033215
  22. 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
  23. Measurement of cytosolic free Ca2+ with quin2.
    Methods Enzymol. 1989;172:230-62 PMID: 2747529
  24. n-Sec1: a neural-specific syntaxin-binding protein.
    Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1445-9 PMID: 8108429
  25. Structure and biological activity of botulinum neurotoxin.
    J Physiol (Paris). 1990;84(3):220-8 PMID: 2074545
  26. Possible role for fatty acyl-coenzyme A in intracellular protein transport.
    Nature. 1987 Mar 19-25;326(6110):309-12 PMID: 3821906
  27. SNAPs, a family of NSF attachment proteins involved in intracellular membrane fusion in animals and yeast.
    Cell. 1990 May 18;61(4):709-21 PMID: 2111733
  28. Implications of the SNARE hypothesis for intracellular membrane topology and dynamics.
    Curr Biol. 1994 Mar 1;4(3):220-33 PMID: 7922327
  29. Membrane protein trafficking through the common apical endosome compartment of polarized Caco-2 cells.
    Mol Biol Cell. 1995 May;6(5):597-610 PMID: 7545032
  30. Receptor-mediated transcytosis of IgA in MDCK cells is via apical recycling endosomes.
    J Cell Biol. 1994 Apr;125(1):67-86 PMID: 8138576
  31. Cellubrevin is a ubiquitous tetanus-toxin substrate homologous to a putative synaptic vesicle fusion protein.
    Nature. 1993 Jul 22;364(6435):346-9 PMID: 8332193
  32. Enigma variations: protein mediators of membrane fusion.
    Cell. 1995 Sep 22;82(6):869-72 PMID: 7553845
  33. Isolated light chains of botulinum neurotoxins inhibit exocytosis. Studies in digitonin-permeabilized chromaffin cells.
    J Biol Chem. 1989 Jun 25;264(18):10354-60 PMID: 2732226
  34. Botulinum neurotoxin light chain inhibits norepinephrine secretion in PC12 cells at an intracellular membranous or cytoskeletal site.
    J Neurochem. 1991 Oct;57(4):1413-21 PMID: 1895112
  35. Effect of nocodazole on vesicular traffic to the apical and basolateral surfaces of polarized MDCK cells.
    J Cell Biol. 1990 Dec;111(6 Pt 1):2365-73 PMID: 2277063
  36. Regulation of protein traffic in polarized epithelial cells.
    Bioessays. 1995 Feb;17(2):129-38 PMID: 7748163
  37. Basolateral to apical transcytosis in polarized cells is indirect and involves BFA and trimeric G protein sensitive passage through the apical endosome.
    J Cell Biol. 1994 Jan;124(1-2):83-100 PMID: 7905002
  38. The synaptic vesicle cycle: a cascade of protein-protein interactions.
    Nature. 1995 Jun 22;375(6533):645-53 PMID: 7791897
  39. Endocytic pathways in polarized Caco-2 cells: identification of an endosomal compartment accessible from both apical and basolateral surfaces.
    J Cell Biol. 1990 Feb;110(2):337-48 PMID: 2298809
  40. Control of protein traffic between distinct plasma membrane domains. Requirement for a novel 108,000 protein in the fusion of transcytotic vesicles with the apical plasma membrane.
    J Biol Chem. 1993 Jan 25;268(3):1876-85 PMID: 8420962
  41. An NSF-like ATPase, p97, and NSF mediate cisternal regrowth from mitotic Golgi fragments.
    Cell. 1995 Sep 22;82(6):905-14 PMID: 7553851
  42. Botulinum neurotoxins serotypes A and E cleave SNAP-25 at distinct COOH-terminal peptide bonds.
    FEBS Lett. 1993 Nov 29;335(1):99-103 PMID: 8243676
  43. Phorbol myristate acetate-mediated stimulation of transcytosis and apical recycling in MDCK cells.
    J Cell Biol. 1994 Mar;124(5):717-27 PMID: 8120094
  44. Postendocytotic sorting of the ligand for the polymeric immunoglobulin receptor in Madin-Darby canine kidney cells.
    J Cell Biol. 1989 Aug;109(2):475-86 PMID: 2760105
  45. The t-SNAREs syntaxin 1 and SNAP-25 are present on organelles that participate in synaptic vesicle recycling.
    J Cell Biol. 1995 Feb;128(4):637-45 PMID: 7860636
  46. An autonomous signal for basolateral sorting in the cytoplasmic domain of the polymeric immunoglobulin receptor.
    Cell. 1991 Jul 12;66(1):65-75 PMID: 2070419
  47. Cell surface polarity in epithelia.
    Annu Rev Cell Biol. 1985;1:243-88 PMID: 3939606
  48. NSF-independent fusion mechanisms.
    Cell. 1995 May 19;81(4):475-7 PMID: 7758102
  49. Molecular mechanisms of clostridial neurotoxins.
    Ann N Y Acad Sci. 1994 Sep 15;733:245-55 PMID: 7978874
  50. Transcytosis of the polymeric immunoglobulin receptor is regulated in multiple intracellular compartments.
    J Biol Chem. 1994 Nov 25;269(47):29474-80 PMID: 7961930
  51. Vesicular transport between the endoplasmic reticulum and the Golgi stack requires the NEM-sensitive fusion protein.
    Nature. 1989 Jun 1;339(6223):397-8 PMID: 2542798
  52. Nerve growth factor induces sensitivity to botulinum neurotoxin type A in norepinephrine-secreting PC12 cells.
    Neurosci Lett. 1993 Dec 24;164(1-2):93-6 PMID: 8152624
  53. Membrane fusion and the cell cycle: Cdc48p participates in the fusion of ER membranes.
    Cell. 1995 Sep 22;82(6):885-93 PMID: 7553849
  54. The molecular machinery for secretion is conserved from yeast to neurons.
    Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2559-63 PMID: 8096639
  55. SNAP family of NSF attachment proteins includes a brain-specific isoform.
    Nature. 1993 Mar 25;362(6418):353-5 PMID: 8455721
  56. Possible role of both the alpha and beta gamma subunits of the heterotrimeric G protein, Gs, in transcytosis of the polymeric immunoglobulin receptor.
    J Biol Chem. 1993 Dec 5;268(34):25824-35 PMID: 8245019
  57. Fusion of sequentially internalized vesicles in alveolar macrophages.
    J Cell Biol. 1990 Apr;110(4):1013-22 PMID: 1691186
  58. Rab GTPases in vesicular transport.
    Curr Opin Cell Biol. 1993 Aug;5(4):613-20 PMID: 8257602
  59. Shift of equilibrium density induced by 3,3'-diaminobenzidine cytochemistry: a new procedure for the analysis and purification of peroxidase-containing organelles.
    J Cell Biol. 1984 Mar;98(3):870-6 PMID: 6699089
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1996-04-01
Pages
1471-81
Language
English
Region
England
NLM ID
8208664
PMCID
PMC450054
Subset
IM
Grants
NINDS NIH HHS · NS17742 · United States
NIAID NIH HHS · R01AI25144 · United States
External Links
PubMed source PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2