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Entry of Challenge Virus Standard (CVS) -11 into N2a cells via a clathrin-mediated, cholesterol-, dynamin-, pH-dependent endocytic pathway.

BACKGROUNDRabies virus (RABV), a member of Lyssavirus of Rhabdoviridae family, is a kind of negative-strand RNA virus. The zoonosis caused by RABV leads to high mortality in animals and humans. Though with the extensive investigation, the mechanisms of RABV entry into cells have not been well charac... Full description

Journal Title: Virology journal June 13, 2019, Vol.16(1), p.80
Main Author: Gao, Jie
Other Authors: Wang, Xinyu , Zhao, Mingxin , Liu, Enhua , Duan, Ming , Guan, Zhenhong , Guo, Yidi , Zhang, Maolin
Format: Electronic Article Electronic Article
Language: English
Subjects:
N2a
ID: E-ISSN: 1743-422X ; DOI: 1743-422X ; DOI: 10.1186/s12985-019-1186-9
Link: http://search.proquest.com/docview/2242816229/?pq-origsite=primo
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title: Entry of Challenge Virus Standard (CVS) -11 into N2a cells via a clathrin-mediated, cholesterol-, dynamin-, pH-dependent endocytic pathway.
format: Article
creator:
  • Gao, Jie
  • Wang, Xinyu
  • Zhao, Mingxin
  • Liu, Enhua
  • Duan, Ming
  • Guan, Zhenhong
  • Guo, Yidi
  • Zhang, Maolin
subjects:
  • Cell Line–Pharmacology
  • Chlorpromazine–Metabolism
  • Cholesterol–Metabolism
  • Clathrin–Metabolism
  • Dynamins–Genetics
  • Endocytosis–Drug Effects
  • Hydrogen-Ion Concentration–Physiology
  • Nucleocapsid Proteins–Physiology
  • RNA Interference–Physiology
  • Rabies Virus–Physiology
  • Virus Internalization–Physiology
  • Clathrin
  • Nucleocapsid Proteins
  • Cholesterol
  • Dynamins
  • Chlorpromazine
  • Caveolin-1
  • Clathrin
  • Endocytosis
  • N2a
  • Rabv
ispartof: Virology journal, June 13, 2019, Vol.16(1), p.80
description: BACKGROUNDRabies virus (RABV), a member of Lyssavirus of Rhabdoviridae family, is a kind of negative-strand RNA virus. The zoonosis caused by RABV leads to high mortality in animals and humans. Though with the extensive investigation, the mechanisms of RABV entry into cells have not been well characterized. METHODSChemical inhibitors and RNA interference (RNAi) were used to analysis RABV internalization pathway. The expression level of viral N protein was examined by quantitative PCR and western blot, and the virus infection in the cells was visualized by fluorescence microscopy. RESULTSWe firstly examined the endocytosis pathway of the challenge virus standard (CVS) -11 strain in N2a cells. Chlorpromazine treatment and knockdown of clathrin heavy chain (CHC) significantly reduced viral entry, which proved clathrin was required. Meanwhile neither nystatin nor knocking down caveolin-1 (Cav1) in N2a cells had an effect on CVS-11 infection, suggesting that caveolae was independent for CVS-11 internalization. And when cholesterol of cell membrane was extracted by MβCD, viral infection was strongly impacted. Additionally by using the specific inhibitor dynasore and ammonium chloride, we verified that dynamin and a low-pH environment were crucial for RABV infection, which was confirmed by confocal microscopy. CONCLUSIONSOur results demonstrated that CVS-11 entered N2a cells through a clathrin-mediated, cholesterol-, pH-, dynamin-required, and caveolae-independent endocytic pathway.
language: eng
source:
identifier: E-ISSN: 1743-422X ; DOI: 1743-422X ; DOI: 10.1186/s12985-019-1186-9
fulltext: fulltext
issn:
  • 1743422X
  • 1743-422X
url: Link


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titleEntry of Challenge Virus Standard (CVS) -11 into N2a cells via a clathrin-mediated, cholesterol-, dynamin-, pH-dependent endocytic pathway.
creatorGao, Jie ; Wang, Xinyu ; Zhao, Mingxin ; Liu, Enhua ; Duan, Ming ; Guan, Zhenhong ; Guo, Yidi ; Zhang, Maolin
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ispartofVirology journal, June 13, 2019, Vol.16(1), p.80
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subjectCell Line–Pharmacology ; Chlorpromazine–Metabolism ; Cholesterol–Metabolism ; Clathrin–Metabolism ; Dynamins–Genetics ; Endocytosis–Drug Effects ; Hydrogen-Ion Concentration–Physiology ; Nucleocapsid Proteins–Physiology ; RNA Interference–Physiology ; Rabies Virus–Physiology ; Virus Internalization–Physiology ; Clathrin ; Nucleocapsid Proteins ; Cholesterol ; Dynamins ; Chlorpromazine ; Caveolin-1 ; Clathrin ; Endocytosis ; N2a ; Rabv
descriptionBACKGROUNDRabies virus (RABV), a member of Lyssavirus of Rhabdoviridae family, is a kind of negative-strand RNA virus. The zoonosis caused by RABV leads to high mortality in animals and humans. Though with the extensive investigation, the mechanisms of RABV entry into cells have not been well characterized. METHODSChemical inhibitors and RNA interference (RNAi) were used to analysis RABV internalization pathway. The expression level of viral N protein was examined by quantitative PCR and western blot, and the virus infection in the cells was visualized by fluorescence microscopy. RESULTSWe firstly examined the endocytosis pathway of the challenge virus standard (CVS) -11 strain in N2a cells. Chlorpromazine treatment and knockdown of clathrin heavy chain (CHC) significantly reduced viral entry, which proved clathrin was required. Meanwhile neither nystatin nor knocking down caveolin-1 (Cav1) in N2a cells had an effect on CVS-11 infection, suggesting that caveolae was independent for CVS-11 internalization. And when cholesterol of cell membrane was extracted by MβCD, viral infection was strongly impacted. Additionally by using the specific inhibitor dynasore and ammonium chloride, we verified that dynamin and a low-pH environment were crucial for RABV infection, which was confirmed by confocal microscopy. CONCLUSIONSOur results demonstrated that CVS-11 entered N2a cells through a clathrin-mediated, cholesterol-, pH-, dynamin-required, and caveolae-independent endocytic pathway.
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titleEntry of Challenge Virus Standard (CVS) -11 into N2a cells via a clathrin-mediated, cholesterol-, dynamin-, pH-dependent endocytic pathway.
descriptionBACKGROUNDRabies virus (RABV), a member of Lyssavirus of Rhabdoviridae family, is a kind of negative-strand RNA virus. The zoonosis caused by RABV leads to high mortality in animals and humans. Though with the extensive investigation, the mechanisms of RABV entry into cells have not been well characterized. METHODSChemical inhibitors and RNA interference (RNAi) were used to analysis RABV internalization pathway. The expression level of viral N protein was examined by quantitative PCR and western blot, and the virus infection in the cells was visualized by fluorescence microscopy. RESULTSWe firstly examined the endocytosis pathway of the challenge virus standard (CVS) -11 strain in N2a cells. Chlorpromazine treatment and knockdown of clathrin heavy chain (CHC) significantly reduced viral entry, which proved clathrin was required. Meanwhile neither nystatin nor knocking down caveolin-1 (Cav1) in N2a cells had an effect on CVS-11 infection, suggesting that caveolae was independent for CVS-11 internalization. And when cholesterol of cell membrane was extracted by MβCD, viral infection was strongly impacted. Additionally by using the specific inhibitor dynasore and ammonium chloride, we verified that dynamin and a low-pH environment were crucial for RABV infection, which was confirmed by confocal microscopy. CONCLUSIONSOur results demonstrated that CVS-11 entered N2a cells through a clathrin-mediated, cholesterol-, pH-, dynamin-required, and caveolae-independent endocytic pathway.
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titleEntry of Challenge Virus Standard (CVS) -11 into N2a cells via a clathrin-mediated, cholesterol-, dynamin-, pH-dependent endocytic pathway.
authorGao, Jie ; Wang, Xinyu ; Zhao, Mingxin ; Liu, Enhua ; Duan, Ming ; Guan, Zhenhong ; Guo, Yidi ; Zhang, Maolin
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abstractBACKGROUNDRabies virus (RABV), a member of Lyssavirus of Rhabdoviridae family, is a kind of negative-strand RNA virus. The zoonosis caused by RABV leads to high mortality in animals and humans. Though with the extensive investigation, the mechanisms of RABV entry into cells have not been well characterized. METHODSChemical inhibitors and RNA interference (RNAi) were used to analysis RABV internalization pathway. The expression level of viral N protein was examined by quantitative PCR and western blot, and the virus infection in the cells was visualized by fluorescence microscopy. RESULTSWe firstly examined the endocytosis pathway of the challenge virus standard (CVS) -11 strain in N2a cells. Chlorpromazine treatment and knockdown of clathrin heavy chain (CHC) significantly reduced viral entry, which proved clathrin was required. Meanwhile neither nystatin nor knocking down caveolin-1 (Cav1) in N2a cells had an effect on CVS-11 infection, suggesting that caveolae was independent for CVS-11 internalization. And when cholesterol of cell membrane was extracted by MβCD, viral infection was strongly impacted. Additionally by using the specific inhibitor dynasore and ammonium chloride, we verified that dynamin and a low-pH environment were crucial for RABV infection, which was confirmed by confocal microscopy. CONCLUSIONSOur results demonstrated that CVS-11 entered N2a cells through a clathrin-mediated, cholesterol-, pH-, dynamin-required, and caveolae-independent endocytic pathway.
doi10.1186/s12985-019-1186-9
urlhttp://search.proquest.com/docview/2242816229/
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date2019-06-13