Early Stages of Influenza Virus Entry into Mv-1 Lung Cells: Involvement of Dynamin

Roy, Ann-Marie; Parker, John S. L.; Parrish, Colin R.; Whittaker, Gary R.

doi:10.1006/viro.1999.0109

Cited by 57 publications

(40 citation statements)

References 55 publications

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“…When expressed at high levels, dominant-negative mutant versions of cellular proteins act by overwhelming the wild-type protein, perturbing its function. Overexpression of Dyn-2K44A or Cav-1 deletion mutants, usually over a short time, has been successfully used to dissect the entry pathways of a plethora of viruses (3,11,28). However, this too is problematic to achieve in the HBV infectivity systems currently available, since primary hepatocytes are notoriously difficult to transfect and the HepaRG cells proved refractory to transient transfections or viral infection/transduction once differentiated (data not shown).…”

Section: Discussionmentioning

confidence: 99%

Hepatitis B Virus Requires Intact Caveolin-1 Function for Productive Infection in HepaRG Cells

Macovei

Radulescu

Lazar

et al. 2010

J Virol

108

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Investigation of the entry pathways of hepatitis B virus (HBV), a member of the family Hepadnaviridae, has been hampered by the lack of versatile in vitro infectivity models. Most concepts of hepadnaviral infection come from the more robust duck HBV system; however, whether the two viruses use the same mechanisms to invade target cells is still a matter of controversy. In this study, we investigate the role of an important plasma membrane component, caveolin-1 (Cav-1), in HBV infection. Caveolins are the main structural components of caveolae, plasma membrane microdomains enriched in cholesterol and sphingolipids, which are involved in the endocytosis of numerous ligands and complex signaling pathways within the cell. We used the HepaRG cell line permissive for HBV infection to stably express dominant-negative Cav-1 and dynamin-2, a GTPase involved in vesicle formation at the plasma membrane and other organelles. The endocytic properties of the newly established cell lines, designated HepaRG Cav-1 , HepaRG Cav-1⌬1-81 , HepaRG Dyn-2 , and HepaRG Dyn-2K44A , were validated using specific markers for different entry routes. The cells maintained their properties during cell culture, supported differentiation, and were permissive for HBV infection. The levels of both HBV transcripts and antigens were significantly decreased in cells expressing the mutant proteins, while viral replication was not directly affected. Chemical inhibitors that specifically inhibit clathrin-mediated endocytosis had no effect on HBV infection. We concluded that HBV requires a Cav-1-mediated entry pathway to initiate productive infection in HepaRG cells.

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Section: Discussionmentioning

confidence: 99%

Hepatitis B Virus Requires Intact Caveolin-1 Function for Productive Infection in HepaRG Cells

Macovei

Radulescu

Lazar

et al. 2010

J Virol

108

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“…It is not known whether this pathway depends on dynamin (Sieczkarski & Whittaker, 2002b). Influenza virus entry in Mv-1 lung cells has been shown to depend on dynamin but further characterization has not been performed (Roy et al, 2000). As the route of entry depends on the host cell type, it is impossible to predict whether influenza virus entry into HeLa cells will also depend on dynamin.…”

Section: Discussionmentioning

confidence: 99%

Clathrin- and caveolae-independent entry of feline infectious peritonitis virus in monocytes depends on dynamin

Hamme

Dewerchin

Cornelissen

et al. 2008

Journal of General Virology

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Feline infectious peritonitis virus (FIPV), a coronavirus that causes a lethal chronic disease in cats, enters feline monocytes via endocytosis. In this study, the pathway of internalization is characterized by evaluating the effect of chemical inhibitors and/or expression of dominantnegative (DN) proteins on the percentage of internalized virions per cell and infection. Further, colocalization studies were performed to determine the involvement of certain cellular internalization proteins. FIPV is not internalized through a clathrin-mediated pathway, as chlorpromazine, amantadine and DN eps15 did not influence virus uptake and FIPV did not co-localize with clathrin. The caveolae-mediated pathway could be excluded based on the inability of genistein and DN caveolin-1 to inhibit virus uptake and lack of co-localization between FIPV and caveolin-1. Dynamin inhibitory peptide and DN dynamin effectively inhibited virus internalization. The inhibitor strongly reduced uptake to 20.3±1.1 % of uptake in untreated cells. In the presence of DN dynamin, uptake was 58.7±3.9 % relative to uptake in untransduced cells. Internalization of FIPV was slightly reduced to 85.0±1.4 and 87.4±6.1 % of internalization in control cells by the sterolbinding drugs nystatin and methyl-b-cyclodextrin, respectively. Rho GTPases were inhibited by Clostridium difficile toxin B, but no effect was observed. These results were confirmed with infection studies showing that infection was not influenced by chlorpromazine, amantadine and genistein, but was significantly reduced by dynamin inhibition and nystatin. In conclusion, these results indicate that FIPV enters monocytes through a clathrin-and caveolae-independent pathway that strongly depends on dynamin and is slightly sensitive to cholesterol depletion. INTRODUCTIONViral entry occurs through a number of successive steps allowing the virus to bring its genome inside the cell. The virus either fuses with the host cell membrane or utilizes the endocytosis machinery of the cell. Endocytosis can be divided in two categories: phagocytosis of large cargo and pinocytosis of smaller cargo (Conner & Schmid, 2003). Phagocytosis is an active and highly regulated process involving specific cell surface receptors and signalling cascades mediated by Rho GTPases (Hall & Nobes, 2000;Conner & Schmid, 2003). Pinocytosis can occur through several pathways. The cargo and its receptor determine which pinocytic pathway will be used (Conner & Schmid, 2003). A first possible route for pinocytosis is 'macropinocytosis' (Swanson & Watts, 1995). Other pathways are named after the main structural compound involved: 'clathrin-mediated endocytosis ' (Brodsky et al., 2001) and 'caveolae-mediated endocytosis' (Pelkmans & Helenius, 2002). Finally, there is a group called 'clathrin-and caveolae-independent pathways'. They are differentiated based on their dependency on cholesterol and/or association with lipid rafts, dynamin and Rho GTPases (Nichols & Lippincott-Schwartz, 2001;Conner & Schmid, 2003). Viruses are specialized in...

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“…HA binding triggers clathrin-dependent receptor-mediated endocytosis of the bound virion through the adapter protein Epsin-1 in a dynamindependent process (7,45,50). Following endocytosis, the virus is trafficked through the endosomal maturation pathway until endosomal acidification triggers the low-pH activation of the HA molecule.…”

mentioning

confidence: 99%

Filamentous Influenza Virus Enters Cells via Macropinocytosis

Rossman

Leser

Lamb

2012

J Virol

127

119

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Influenza virus is pleiomorphic, producing both spherical (100-nm-diameter) and filamentous (100-nm by 20-m) virions. While the spherical virions are known to enter host cells through exploitation of clathrin-mediated endocytosis, the entry pathway for filamentous virions has not been determined, though the existence of an alternative, non-clathrin-, non-caveolin-mediated entry pathway for influenza virus has been known for many years. In this study, we confirm recent results showing that influenza virus utilizes macropinocytosis as an alternate entry pathway. Furthermore, we find that filamentous influenza viruses use macropinocytosis as the primary entry mechanism. Virions enter cells as intact filaments within macropinosomes and are trafficked to the acidic late-endosomal compartment. Low pH triggers a conformational change in the M2 ion channel protein, altering membrane curvature and leading to a fragmentation of the filamentous virions. This fragmentation may enable moreefficient fusion between the viral and endosomal membranes.

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Early Stages of Influenza Virus Entry into Mv-1 Lung Cells: Involvement of Dynamin

Cited by 57 publications

References 55 publications

Hepatitis B Virus Requires Intact Caveolin-1 Function for Productive Infection in HepaRG Cells

Hepatitis B Virus Requires Intact Caveolin-1 Function for Productive Infection in HepaRG Cells

Clathrin- and caveolae-independent entry of feline infectious peritonitis virus in monocytes depends on dynamin

Filamentous Influenza Virus Enters Cells via Macropinocytosis

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