Entry of Feline Calicivirus Is Dependent on Clathrin-Mediated Endocytosis and Acidification in Endosomes

Stuart, Amanda D.; Brown, T. D. K.

doi:10.1128/jvi.02452-05

Cited by 89 publications

(103 citation statements)

References 65 publications

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“…In this study, we have further examined the entry route used by MNV-1 to infect RAW264.7 cells. Initially, we investigated if infection by MNV-1 is dependent on CME, since FCV (which is closely related to MNV-1) is internalized via this pathway (Stuart & Brown, 2006). Our experiments show that DN Eps-15 and AP-2 knockdown inhibited transferrin uptake but had no Fig.…”

Section: Discussionmentioning

confidence: 85%

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Murine norovirus-1 cell entry is mediated through a non-clathrin-, non-caveolae-, dynamin- and cholesterol-dependent pathway

Gerondopoulos

Jackson

Monaghan

et al. 2010

Journal of General Virology

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For many viruses, endocytosis and exposure to the low pH within acidic endosomes is essential for infection. It has previously been reported that feline calicivirus uses clathrin-mediated endocytosis for entry into mammalian cells. Here, we report that infection of RAW264.7 macrophages by the closely related murine norovirus-1 (MNV-1) does not require the clathrin pathway, as infection was not inhibited by expression of dominant-negative Eps15 or by knockdown of the adaptin-2 complex. Further, infection was not inhibited by reagents that raise endosomal pH. RAW264.7 macrophages were shown not to express caveolin, and flotillin depletion did not inhibit infection, suggesting that caveolae and the flotillin pathway are not required for cell entry. However, MNV-1 infection was inhibited by methyl-b-cyclodextrin and the dynamin inhibitor, dynasore. Addition of these drugs to the cells after a period of virus internalization did not inhibit infection, suggesting the involvement of cholesterol-sensitive lipid rafts and dynamin in the entry mechanism. Macropinocytosis (MPC) was shown to be active in RAW264.7 macrophages (as indicated by uptake of dextran) and could be blocked by 5-(N-ethyl-N-isopropyl) amiloride (EIPA), which is reported to inhibit this pathway. However, infection was enhanced in the presence of EIPA. Similarly, actin disruption, which also inhibits MPC, resulted in enhanced infection. These results suggest that MPC could contribute to virus degradation or that inhibition of MPC could lead to the upregulation of other endocytic pathways of virus uptake. INTRODUCTIONThe family Caliciviridae is divided into four genera: Vesivirus, Lagovirus, Sapovirus and Norovirus. The human noroviruses are the most common cause of acute viral gastroenteritis, especially in industrialized countries (Lopman et al., 2003); there is currently no treatment or vaccine for these viruses. In addition, there is still no routine tissue culture system for the propagation of human noroviruses, but the recent discovery of murine norovirus-1 (MNV-1) has provided an efficient model system for the study of norovirus replication, as this virus replicates efficiently in murine macrophages and dendritic cells (DCs) (Karst et al., 2003;Wobus et al., 2004). MNV-1 is highly prevalent in laboratory mice and has been shown to be lethal to mice with impaired innate immunity (Hsu et al., 2006;Karst et al., 2003;Muller et al., 2007).Noroviruses are non-enveloped and contain a singlestranded RNA genome of about 7.3 kb, which is linked to the viral VPg protein at the 59 end and is polyadenylated at the 39 end (Karst et al., 2003;Wobus et al., 2004). The genome is organized into four open reading frames (ORF-1-4). ORF-1 encodes the non-structural proteins, whereas ORF-2 and ORF-3 encode structural proteins (Sosnovtsev et al., 2006). ORF-4 was recently identified within the MNV-1 genome and encodes a single protein of unknown function (Thackray et al., 2007).For many viruses, infection requires virus uptake by endocytosis. A number of different endo...

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Section: Discussionmentioning

confidence: 85%

“…In addition, FCV has been shown to bind to 2,6-sialic acid (Stuart & Brown, 2007), which could serve as a co-receptor for infection. Also, FCV is known to be internalized by CME as infection is inhibited by expression of dominant-negative (DN) mutants of proteins (Eps15 and Rab5) that are normally involved in CME (Stuart & Brown, 2006).…”

Section: Introductionmentioning

confidence: 99%

Murine norovirus-1 cell entry is mediated through a non-clathrin-, non-caveolae-, dynamin- and cholesterol-dependent pathway

Gerondopoulos

Jackson

Monaghan

et al. 2010

Journal of General Virology

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“…Thus, we tested whether the depletion of clathrin and its adaptor proteins affect BPV entry into the cells. Chlorpromazine (CHPZ) prevents assembly and disassembly of clathrin lattices at the cell surface and on endosomes, that is, it prevents assembly of coated pits (Blanchard et al, 2006;Brindley & Maury, 2008;Stuart & Brown, 2006). CHPZ was maintained in the cultures for the duration of BPV infection.…”

Section: Bpv Entry Into Cells Utilizes Clathrin and Endosomal Acidifimentioning

confidence: 99%

Bovine parvovirus uses clathrin-mediated endocytosis for cell entry

Dudleenamjil¹,

Lin²,

Dredge³

et al. 2010

Journal of General Virology

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Entry events of bovine parvovirus (BPV) were studied. Transmission electron micrographs of infected cells showed virus particles in cytoplasmic vesicles. Chemical inhibitors that block certain aspects of the cellular machinery were employed to assess viral dependency upon those cellular processes. Chlorpromazine, ammonium chloride, chloroquine and bafilamicin A1 were used to inhibit acidification of endosomes and clathrin-associated endocytosis. Nystatin was used as an inhibitor of the caveolae pathway. Cytochalasin D and ML-7 were used to inhibit actin and myosin functions, respectively. Nocodazole and colchicine were employed to inhibit microtubule activity. Virus entry was assessed by measuring viral transcription using real-time PCR, synthesis of capsid protein and assembly of infectious progeny virus in the presence of inhibitor blockage. The results indicated that BPV entry into embryonic bovine trachael cells utilizes endocytosis in clathrin-coated vesicles, is dependent upon acidification, and appears to be associated with actin and microtubule dependency. Evidence for viral entry through caveolae was not obtained. These findings provide a fuller understanding of the early cell-entry events of the replication cycle for members of the genus Bocavirus.

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“…Several possible pathways of receptor-mediated endocytosis are recognized: clathrin-and caveolae-mediated endocytosis, macropinocytosis, and novel nonclathrin/noncaveolae pathways (Brindley & Maury, 2008;Damm et al, 2005;Dimitrov, 2004;Kee et al, 2004;Marsh & Helenius, 2006;Meier & Greber, 2004;Meier et al, 2002;Mercer & Helenius, 2009;Pelkmans & Helenius, 2003;Pelkmans et al 2004;Sieczkarski & Whittaker, 2002;Stuart & Brown, 2006). To date, for viral infections that have been studied, viruses mostly take advantage of clathrin-mediated endocytosis for internalization (Marsh & Helenius, 2006;Pelkmans & Helenius, 2003;Sieczkarski & Whittaker, 2002).…”

Section: Engulfmentmentioning

confidence: 99%

“…It has been widely used as a probe to study acid-mediated viral trafficking using a variety of virus models. Entry pathway studies of mouse polyomavirus (Liebl et al, 2006), feline calicivirus (Stuart & Brown, 2006), bovine viral diarrhea virus (Lecot et al, 2005), AAV (Bartlett et al, 2000), MVM (Mani et al, 2006;Ros et al, 2002), mouse hepatitis virus type 2 (Pu & Zhang, 2008), poliovirus , baculovirus (Long et al, 2006), hepatitis C virus (Blanchard et al, 2006), influenza virus (Guinea & Carrasco, 1995;Sieczkarski et al, 2003), HIV-1 (Fredericksen et al, 2002), and human rhinovirus 14 (Bayer et al, 1999) have analyzed the effects of Baf A1 on virus trafficking and examined the role of endosomal acidification in a route that leads to viral replication. For studies of parvovirus entry Baf A1 may be an inhibitor preferred over chlorpromazine because chlorpromazine is not only an inhibitor of clathrin lattice processing, but is also an inhibitor of phospholipase A2.…”

Section: Engulfmentmentioning

confidence: 99%