The coronavirus transmissible gastroenteritis virus causes infection after receptor-mediated endocytosis and acid-dependent fusion with an intracellular compartment

Gert Helge Hansen; B Delmas; L Besnardeau; L K Vogel; H Laude; H Sjöström; Ove Norén

The coronavirus transmissible gastroenteritis virus causes infection after receptor-mediated endocytosis and acid-dependent fusion with an intracellular compartment

Gert Helge Hansen, B Delmas, L Besnardeau, L K Vogel, H Laude, H Sjöström, Ove Norén

Institut for Cellulær og Molekylær Medicin

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

52 Citationer (Scopus)

Abstract

Udgivelsesdato: 1998-Jan

Originalsprog	Engelsk
Tidsskrift	Journal of Virology
Vol/bind	72
Udgave nummer	1
Sider (fra-til)	527-34
Antal sider	7
ISSN	0022-538X
Status	Udgivet - 1998

Bibliografisk note

Keywords: Ammonium Chloride; Animals; Anti-Bacterial Agents; Antigens, CD13; Cell Compartmentation; Cell Line; Cell Membrane; Dogs; Endocytosis; Enzyme Inhibitors; Gastroenteritis, Transmissible, of Swine; Hydrogen-Ion Concentration; Lysosomes; Macrolides; Membrane Fusion; Microscopy, Electron; Proton Pumps; Receptors, Cell Surface; Swine; Transmissible gastroenteritis virus; Virus Replication

Citationsformater

@article{4e1b4440e30e11ddb5fc000ea68e967b,

title = "The coronavirus transmissible gastroenteritis virus causes infection after receptor-mediated endocytosis and acid-dependent fusion with an intracellular compartment",

abstract = "Aminopeptidase N is a species-specific receptor for transmissible gastroenteritis virus (TGEV), which infects piglets, and for the 229E virus, which infects humans. It is not known whether these coronaviruses are endocytosed before fusion with a membrane of the target cell, causing a productive infection, or whether they fuse directly with the plasma membrane. We have studied the interaction between TGEV and a cell line (MDCK) stably expressing recombinant pig aminopeptidase N (pAPN). By electron microscopy and flow cytometry, TGEV was found to be associated with the plasma membrane after adsorption to the pAPN-MDCK cells. TGEV was also observed in endocytic pits and apical vesicles after 3 to 10 min of incubation at 38 degrees C. The number of pits and apical vesicles was increased by the TGEV incubation, indicating an increase in endocytosis. After 10 min of incubation, a distinct TGEV-pAPN-containing population of large intracellular vesicles, morphologically compatible with endosomes, was found. A higher density of pAPN receptors was observed in the pits beneath the virus particles than in the surrounding plasma membrane, indicating that TGEV recruits pAPN receptors before endocytosis. Ammonium chloride and bafilomycin A1 markedly inhibited the TGEV infection as judged from virus production and protein biosynthesis analyses but did so only when added early in the course of the infection, i.e., about 1 h after the start of endocytosis. Together our results point to an acid intracellular compartment as the site of fusion for TGEV.",

author = "Hansen, {Gert Helge} and B Delmas and L Besnardeau and Vogel, {L K} and H Laude and H Sj{\"o}str{\"o}m and Ove Nor{\'e}n",

note = "Keywords: Ammonium Chloride; Animals; Anti-Bacterial Agents; Antigens, CD13; Cell Compartmentation; Cell Line; Cell Membrane; Dogs; Endocytosis; Enzyme Inhibitors; Gastroenteritis, Transmissible, of Swine; Hydrogen-Ion Concentration; Lysosomes; Macrolides; Membrane Fusion; Microscopy, Electron; Proton Pumps; Receptors, Cell Surface; Swine; Transmissible gastroenteritis virus; Virus Replication",

year = "1998",

language = "English",

volume = "72",

pages = "527--34",

journal = "Journal of Virology",

issn = "0022-538X",

publisher = "American Society for Microbiology",

number = "1",

}

TY - JOUR

T1 - The coronavirus transmissible gastroenteritis virus causes infection after receptor-mediated endocytosis and acid-dependent fusion with an intracellular compartment

AU - Hansen, Gert Helge

AU - Delmas, B

AU - Besnardeau, L

AU - Vogel, L K

AU - Laude, H

AU - Sjöström, H

AU - Norén, Ove

N1 - Keywords: Ammonium Chloride; Animals; Anti-Bacterial Agents; Antigens, CD13; Cell Compartmentation; Cell Line; Cell Membrane; Dogs; Endocytosis; Enzyme Inhibitors; Gastroenteritis, Transmissible, of Swine; Hydrogen-Ion Concentration; Lysosomes; Macrolides; Membrane Fusion; Microscopy, Electron; Proton Pumps; Receptors, Cell Surface; Swine; Transmissible gastroenteritis virus; Virus Replication

PY - 1998

Y1 - 1998

N2 - Aminopeptidase N is a species-specific receptor for transmissible gastroenteritis virus (TGEV), which infects piglets, and for the 229E virus, which infects humans. It is not known whether these coronaviruses are endocytosed before fusion with a membrane of the target cell, causing a productive infection, or whether they fuse directly with the plasma membrane. We have studied the interaction between TGEV and a cell line (MDCK) stably expressing recombinant pig aminopeptidase N (pAPN). By electron microscopy and flow cytometry, TGEV was found to be associated with the plasma membrane after adsorption to the pAPN-MDCK cells. TGEV was also observed in endocytic pits and apical vesicles after 3 to 10 min of incubation at 38 degrees C. The number of pits and apical vesicles was increased by the TGEV incubation, indicating an increase in endocytosis. After 10 min of incubation, a distinct TGEV-pAPN-containing population of large intracellular vesicles, morphologically compatible with endosomes, was found. A higher density of pAPN receptors was observed in the pits beneath the virus particles than in the surrounding plasma membrane, indicating that TGEV recruits pAPN receptors before endocytosis. Ammonium chloride and bafilomycin A1 markedly inhibited the TGEV infection as judged from virus production and protein biosynthesis analyses but did so only when added early in the course of the infection, i.e., about 1 h after the start of endocytosis. Together our results point to an acid intracellular compartment as the site of fusion for TGEV.

AB - Aminopeptidase N is a species-specific receptor for transmissible gastroenteritis virus (TGEV), which infects piglets, and for the 229E virus, which infects humans. It is not known whether these coronaviruses are endocytosed before fusion with a membrane of the target cell, causing a productive infection, or whether they fuse directly with the plasma membrane. We have studied the interaction between TGEV and a cell line (MDCK) stably expressing recombinant pig aminopeptidase N (pAPN). By electron microscopy and flow cytometry, TGEV was found to be associated with the plasma membrane after adsorption to the pAPN-MDCK cells. TGEV was also observed in endocytic pits and apical vesicles after 3 to 10 min of incubation at 38 degrees C. The number of pits and apical vesicles was increased by the TGEV incubation, indicating an increase in endocytosis. After 10 min of incubation, a distinct TGEV-pAPN-containing population of large intracellular vesicles, morphologically compatible with endosomes, was found. A higher density of pAPN receptors was observed in the pits beneath the virus particles than in the surrounding plasma membrane, indicating that TGEV recruits pAPN receptors before endocytosis. Ammonium chloride and bafilomycin A1 markedly inhibited the TGEV infection as judged from virus production and protein biosynthesis analyses but did so only when added early in the course of the infection, i.e., about 1 h after the start of endocytosis. Together our results point to an acid intracellular compartment as the site of fusion for TGEV.

M3 - Journal article

C2 - 9420255

SN - 0022-538X

VL - 72

SP - 527

EP - 534

JO - Journal of Virology

JF - Journal of Virology

IS - 1

ER -