scholarly journals Ganglioside GD1a Restores Infectibility to Mouse Cells Lacking Functional Receptors for Polyomavirus

2005 ◽  
Vol 79 (1) ◽  
pp. 615-618 ◽  
Author(s):  
Joanna Gilbert ◽  
Jean Dahl ◽  
Cathy Riney ◽  
John You ◽  
Cunqi Cui ◽  
...  
Keyword(s):  
Simian Virus 40 ◽  
Simian Virus ◽  
Cell Entry ◽  
Heterologous Host ◽  
Virus Binding ◽  
Rat Glioma ◽  
Caveolin 1 ◽  
Ganglioside Biosynthesis ◽  
A Cell ◽  
Mouse Cells

ABSTRACT Recent investigations on the pathway of cell entry by polyomavirus (Py) and simian virus 40 (SV40) have defined specific gangliosides as functional receptors mediating virus binding and transport from the plasma membrane to the endoplasmic reticulum (B. Tsai et al., EMBO J. 22:4346-4355, 2003; Gilbert and Benjamin, in press). These studies were carried out with C6 rat glioma cells, a heterologous host chosen for its known deficiency in ganglioside biosynthesis. Here, a cell genetic approach was undertaken to identify components required for the early steps of infection using mouse cells as the natural host for Py. Receptor-negative (R−) mouse cells, screened based on resistance to Py infection, were shown to bind Py but failed to allow entry of the virus. R− cells were also found to be resistant to SV40. Infectibility was restored or enhanced by the addition of the same specific gangliosides found in earlier studies with C6 cells. In one R− line, overexpression of caveolin-1 also increased infectibility. These results support and extend findings on gangliosides in lipid rafts as functional receptors and mediators of internalization for Py and SV40.

scholarly journals Assembly and trafficking of caveolar domains in the cell

2005 ◽  
Vol 170 (5) ◽  
pp. 769-779 ◽  
Author(s):  
Akiko Tagawa ◽  
Anna Mezzacasa ◽  
Arnold Hayer ◽  
Andrea Longatti ◽  
Lucas Pelkmans ◽  
...  
Keyword(s):  
Total Internal Reflection ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Cell Entry ◽  
Cholesterol Depletion ◽  
Membrane Domains ◽  
Vesicle Traffic ◽  
Caveolin 1 ◽  
Transport Vesicles ◽  
Microscopy Techniques

Using total internal reflection fluorescence microscopy (TIR-FM), fluorescence recovery after photobleaching (FRAP), and other light microscopy techniques, we analyzed the dynamics, the activation, and the assembly of caveolae labeled with fluorescently tagged caveolin-1 (Cav1). We found that when activated by simian virus 40 (SV40), a nonenveloped DNA virus that uses caveolae for cell entry, the fraction of mobile caveolae was dramatically enhanced both in the plasma membrane (PM) and in the caveosome, an intracellular organelle that functions as an intermediate station in caveolar endocytosis. Activation also resulted in increased microtubule (MT)-dependent, long-range movement of caveolar vesicles. We generated heterokaryons that contained GFP- and RFP-tagged caveolae by fusing cells expressing Cav1-GFP and -RFP, respectively, and showed that even when activated, individual caveolar domains underwent little exchange of Cav1. Only when the cells were subjected to transient cholesterol depletion, did the caveolae domain exchange Cav1. Thus, in contrast to clathrin-, or other types of coated transport vesicles, caveolae constitute stable, cholesterol-dependent membrane domains that can serve as fixed containers through vesicle traffic. Finally, we identified the Golgi complex as the site where newly assembled caveolar domains appeared first.

scholarly journals A Raft-derived, Pak1-regulated Entry Participates in α2β1 Integrin-dependent Sorting to Caveosomes

2008 ◽  
Vol 19 (7) ◽  
pp. 2857-2869 ◽  
Author(s):  
Mikko Karjalainen ◽  
Elina Kakkonen ◽  
Paula Upla ◽  
Heli Paloranta ◽  
Pasi Kankaanpää ◽  
...  
Keyword(s):  
Simian Virus 40 ◽  
Fluid Phase ◽  
Simian Virus ◽  
Cell Entry ◽  
Phosphatidylinositol 3 Kinase ◽  
Caveolin 1 ◽  
Entry Pathway ◽  
Integrin Clustering ◽  
P21 Activated Kinase ◽  
Α2β1 Integrin

We have previously shown that a human picornavirus echovirus 1 (EV1) is transported to caveosomes during 2 h together with its receptor α2β1 integrin. Here, we show that the majority of early uptake does not occur through caveolae. α2β1 integrin, clustered by antibodies or by EV1 binding, is initially internalized from lipid rafts into tubulovesicular structures. These vesicles accumulate fluid-phase markers but do not initially colocalize with caveolin-1 or internalized simian virus 40 (SV40). Furthermore, the internalized endosomes do not contain glycosylphosphatidylinositol (GPI)-anchored proteins or flotillin 1, suggesting that clustered α2β1 integrin does not enter the GPI-anchored protein enriched endosomal compartment or flotillin pathways, respectively. Endosomes mature further into larger multivesicular bodies between 15 min to 2 h and concomitantly recruit caveolin-1 or SV40 inside. Cell entry is regulated by p21-activated kinase (Pak)1, Rac1, phosphatidylinositol 3-kinase, phospholipase C, and actin but not by dynamin 2 in SAOS-α2β1 cells. An amiloride analog, 5-(N-ethyl-N-isopropanyl) amiloride, blocks infection, causes integrin accumulation in early tubulovesicular structures, and prevents their structural maturation into multivesicular structures. Our results together suggest that α2β1 integrin clustering defines its own entry pathway that is Pak1 dependent but clathrin and caveolin independent and that is able to sort cargo to caveosomes.

scholarly journals Nonselective expression of simian virus 40 large tumor antigen fragments in mouse cells.

1982 ◽  
Vol 79 (6) ◽  
pp. 2064-2067 ◽  
Author(s):  
V. B. Reddy ◽  
S. S. Tevethia ◽  
M. J. Tevethia ◽  
S. M. Weissman
Keyword(s):  
Tumor Antigen ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Large Tumor ◽  
Large Tumor Antigen ◽  
Mouse Cells

scholarly journals In vivo competition of delta-crystallin gene expression by DNA fragments containing a GC box.

1986 ◽  
Vol 6 (11) ◽  
pp. 4130-4132 ◽  
Author(s):  
S Hayashi ◽  
H Kondoh
Keyword(s):  
Gene Expression ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Dna Fragments ◽  
Specific Expression ◽  
Mouse Cells ◽  
Gc Box ◽  
Simplex Virus

Expression of the chicken delta-crystallin gene 1 injected into the nuclei of mouse cells is lens specific. Coinjection of GC box-containing DNA fragments from delta-crystallin, simian virus 40 early, and herpes simplex virus type 1 tk promoters effectively suppressed delta-crystallin expression in the lens, but coinjection with DNA fragments not containing the GC box did not. This suppression was likely due to the competition of an Sp1-like transcription factor(s) and indicates involvement of the apparently ubiquitous factor(s) in the tissue-specific expression of the delta-crystallin gene.

scholarly journals N-Glycolyl GM1 Ganglioside as a Receptor for Simian Virus 40

2007 ◽  
Vol 81 (23) ◽  
pp. 12846-12858 ◽  
Author(s):  
Maria A. Campanero-Rhodes ◽  
Alicia Smith ◽  
Wengang Chai ◽  
Sandro Sonnino ◽  
Laura Mauri ◽  
...  
Keyword(s):  
Simian Virus 40 ◽  
High Specificity ◽  
Simian Virus ◽  
Receptor Expression ◽  
Host Cells ◽  
Gm1 Ganglioside ◽  
Ganglioside Gm1 ◽  
Virus Binding ◽  
Host Interactions ◽  
Sv40 Infection

ABSTRACT Carbohydrate microarrays have emerged as powerful tools in analyses of microbe-host interactions. Using a microarray with 190 sequence-defined oligosaccharides in the form of natural glycolipids and neoglycolipids representative of diverse mammalian glycans, we examined interactions of simian virus 40 (SV40) with potential carbohydrate receptors. While the results confirmed the high specificity of SV40 for the ganglioside GM1, they also revealed that N-glycolyl GM1 ganglioside [GM1(Gc)], which is characteristic of simian species and many other nonhuman mammals, is a better ligand than the N-acetyl analog [GM1(Ac)] found in mammals, including humans. After supplementing glycolipid-deficient GM95 cells with GM1(Ac) and GM1(Gc) gangliosides and the corresponding neoglycolipids with phosphatidylethanolamine lipid groups, it was found that GM1(Gc) analogs conferred better virus binding and infectivity. Moreover, we visualized the interaction of NeuGc with VP1 protein of SV40 by molecular modeling and identified a conformation for GM1(Gc) ganglioside in complex with the virus VP1 pentamer that is compatible with its presentation as a membrane receptor. Our results open the way not only to detailed studies of SV40 infection in relation to receptor expression in host cells but also to the monitoring of changes that may occur with time in receptor usage by the virus.

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