[31] Fusogenic reconstituted sendai virus envelopes as a vehicle for introducing DNA into viable mammalian cells

1983 ◽  
pp. 492-512 ◽  
Author(s):  
A. Vainstein ◽  
A. Razin ◽  
A. Graessmann ◽  
A. Loyter
Keyword(s):  
Mammalian Cells ◽  
Sendai Virus

scholarly journals Double-Layered Membrane Vesicles Released from Mammalian Cells Infected with Sendai Virus Expressing the Matrix Protein of Vesicular Stomatitis Virus

Virology ◽  
1999 ◽  
Vol 263 (1) ◽  
pp. 230-243 ◽  
Author(s):  
Takemasa Sakaguchi ◽  
Tsuneo Uchiyama ◽  
Yutaka Fujii ◽  
Katsuhiro Kiyotani ◽  
Atsushi Kato ◽  
...  
Keyword(s):  
Vesicular Stomatitis Virus ◽  
Mammalian Cells ◽  
Matrix Protein ◽  
Sendai Virus ◽  
Membrane Vesicles ◽  
The Matrix ◽  
Vesicular Stomatitis

scholarly journals A Role for the Sendai Virus P Protein Trimer in RNA Synthesis

1998 ◽  
Vol 72 (5) ◽  
pp. 4274-4280 ◽  
Author(s):  
Joseph Curran
Keyword(s):  
Amino Acids ◽  
Mammalian Cells ◽  
Essential Role ◽  
Rna Synthesis ◽  
Sendai Virus ◽  
Coiled Coil ◽  
The Third ◽  
P Protein ◽  
P Proteins

ABSTRACT The SeV P protein is found as a homotrimer (P3) when it is expressed in mammalian cells, and trimerization is mediated by a predicted coiled-coil motif which maps within amino acids (aa) 344 to 411 (the BoxA region). The bacterially expressed protein also appears to be trimeric, apparently precluding a role for phosphorylation in the association of the P monomers. I have examined the role of P trimerization both in the protein’s interaction with the nucleocapsid (N:RNA) template and in the protein’s function on the template during RNA synthesis. As with the results of earlier experiments (32), I found that both the BoxA and BoxC (aa 479 to 568) regions were required for stable binding of P to the N:RNA. Binding was also observed with P proteins containing less than three BoxC regions, suggesting that trimerization may be required to permit contacts between multiple BoxC regions and the N:RNA. However, these heterologous trimers failed to function in viral RNA synthesis, indicating that the third C-terminal leg of the trimer plays an essential role in P function on the template. We speculate that this function may involve the movement of P (and possibly the polymerase complex) on the template and the maintenance of processivity.

scholarly journals LGP2 virus sensor enhances apoptosis by upregulating apoptosis regulatory genes through TRBP-bound miRNAs during viral infection

2019 ◽  
Vol 48 (3) ◽  
pp. 1494-1507 ◽  
Author(s):  
Tomoko Takahashi ◽  
Yuko Nakano ◽  
Koji Onomoto ◽  
Mitsutoshi Yoneyama ◽  
Kumiko Ui-Tei
Keyword(s):  
Viral Infection ◽  
Rna Silencing ◽  
Mammalian Cells ◽  
Rna Binding ◽  
Sendai Virus ◽  
Cysteine Proteases ◽  
Regulatory Genes ◽  
Type I ◽  
Member Laboratory ◽  
Sensor Proteins

Abstract During viral infection, viral nucleic acids are detected by virus sensor proteins including toll-like receptor 3 or retinoic acid-inducible gene I-like receptors (RLRs) in mammalian cells. Activation of these virus sensor proteins induces type-I interferon production and represses viral replication. Recently, we reported that an RLR family member, laboratory of genetics and physiology 2 (LGP2), modulates RNA silencing by interacting with an RNA silencing enhancer, TAR-RNA binding protein (TRBP). However, the biological implications remained unclear. Here, we show that LGP2 enhances apoptosis by upregulating apoptosis regulatory genes during viral infection. Sendai virus (SeV) infection increased LGP2 expression approximately 900 times compared to that in non-virus-infected cells. Then, the induced LGP2 interacted with TRBP, resulting in the inhibition of maturation of the TRBP-bound microRNA (miRNA) and its subsequent RNA silencing activity. Gene expression profiling revealed that apoptosis regulatory genes were upregulated during SeV infection: caspases-2, -8, -3 and -7, four cysteine proteases with key roles in apoptosis, were upregulated directly or indirectly through the repression of a typical TRBP-bound miRNA, miR-106b. Our findings may shed light on the mechanism of apoptosis, induced by the TRBP-bound miRNAs through the interaction of TRBP with LGP2, as an antiviral defense system in mammalian cells.

scholarly journals Innate Cellular Response to Virus Particle Entry Requires IRF3 but Not Virus Replication

2004 ◽  
Vol 78 (4) ◽  
pp. 1706-1717 ◽  
Author(s):  
Susan E. Collins ◽  
Ryan S. Noyce ◽  
Karen L. Mossman
Keyword(s):  
Virus Replication ◽  
Mammalian Cells ◽  
Cellular Response ◽  
Sendai Virus ◽  
Antiviral Response ◽  
Interferon Production ◽  
Virus Infections ◽  
Mrna Accumulation ◽  
Enveloped Virus ◽  
Hsv 1

ABSTRACT Mammalian cells respond to virus infections by eliciting both innate and adaptive immune responses. One of the most effective innate antiviral responses is the production of alpha/beta interferon and the subsequent induction of interferon-stimulated genes (ISGs), whose products collectively limit virus replication and spread. Following viral infection, interferon is produced in a biphasic fashion that involves a number of transcription factors, including the interferon regulatory factors (IRFs) 1, 3, 7, and 9. In addition, virus infection has been shown to directly induce ISGs in the absence of prior interferon production through the activation of IRF3. This process is believed to require virus replication and results in IRF3 hyperphosphorylation, nuclear localization, and proteasome-mediated degradation. Previously, we and others demonstrated that herpes simplex virus type 1 (HSV-1) induces ISGs and an antiviral response in fibroblasts in the absence of both interferon production and virus replication. In this report, we show that the entry of enveloped virus particles from diverse virus families elicits a similar innate response. This process requires IRF3, but not IRF1, IRF7, or IRF9. Following virus replication, the large DNA viruses HSV-1 and vaccinia virus effectively inhibit ISG mRNA accumulation, whereas the small RNA viruses Newcastle disease virus, Sendai virus, and vesicular stomatitis virus do not. In addition, we found that IRF3 hyperphosphorylation and degradation do not correlate with ISG and antiviral state induction but instead serve as a hallmark of productive virus replication, particularly following a high-multiplicity infection. Collectively, these data suggest that virus entry triggers an innate antiviral response mediated by IRF3 and that subsequent virus replication results in posttranslational modification of IRF3, such as hyperphosphorylation, depending on the nature of the incoming virus.

scholarly journals The biochemistry of virus-induced cell fusion. Changes in membrane integrity

1974 ◽  
Vol 140 (3) ◽  
pp. 405-411 ◽  
Author(s):  
Charles A. Pasternak ◽  
Kingsley J. Micklem
Keyword(s):  
Cell Fusion ◽  
Viral Entry ◽  
Mammalian Cells ◽  
Sendai Virus ◽  
Membrane Integrity ◽  
Intracellular Metabolites ◽  
Nitrophenyl Phosphate ◽  
Biochemical Studies ◽  
Membrane Components ◽  
Interesting System

1. Phospholipids prelabelled with [14C]acetate, [32P]phosphate, [3H]- or [14C]-choline or [3H]inositol are not significantly degraded during fusion of Lettrée cells mediated by Sendai virus, nor are carbohydrates prelabelled with [3H]fucose, [14C]galactose or [3H]glucosamine. Less than 1nmol of lysophosphatidylcholine/107 cells is formed during fusion. Diethyl p-nitrophenyl phosphate, which inhibits phospholipase A by more than 95% has no effect on fusion. It is concluded that none of the events leading to cell fusion is accompanied by significant turnover of phospholipids or other membrane components. 2. Intracellular K+ leaks out during virally mediated cell fusion; the loss is not as extensive as that of intracellularly accumulated choline or deoxyglucose. Movement of Ca2+ into or out of cells could not be detected. 3. At concentrations of Lettrée cells insufficient to be agglutinated by virus, intracellularly accumulated choline and deoxyglucose leak out. Agglutination caused by concanavalin A does not result in leakage of intracellular metabolites. 4. P815Y cells, which agglutinate but do not fuse in the presence of virus, show leakage of intracellularly accumulated metabolites. The extent of leakage does not alter during the G1 and S periods of the cell cycle. 5. Leakage is inhibited by Ca2+, but is unaffected by EDTA. 6. It is concluded that the interaction of Sendai virus with mammalian cells causes a weakening of membrane integrity so that intracellular metabolites leak out. Such destabilization may facilitate viral entry and is therefore an interesting system for further biochemical studies.

scholarly journals AIP1/Alix Is a Binding Partner of Sendai Virus C Protein and Facilitates Virus Budding

2005 ◽  
Vol 79 (14) ◽  
pp. 8933-8941 ◽  
Author(s):  
Takemasa Sakaguchi ◽  
Atsushi Kato ◽  
Fumihiro Sugahara ◽  
Yukie Shimazu ◽  
Makoto Inoue ◽  
...  
Keyword(s):  
Membrane Trafficking ◽  
Mammalian Cells ◽  
Sendai Virus ◽  
Host Protein ◽  
Dependent Manner ◽  
Virus Budding ◽  
C Protein ◽  
Virus Like Particles ◽  
Infected Cells ◽  
Low Efficiency

ABSTRACT The C protein, an accessory protein of Sendai virus (SeV), has anti-interferon capacity and suppresses viral RNA synthesis. In addition, it is thought that the C protein is involved in virus budding because of the low efficiency of release of progeny virions from C-knockout virus-infected cells and because of the requirement of the C protein for efficient release of virus-like particles. Here, we identified AIP1/Alix, a host protein involved in apoptosis and endosomal membrane trafficking, as an interacting partner of the C protein using a yeast two-hybrid system. The amino terminus of AIP1/Alix and the carboxyl terminus of the C protein are important for the interaction in mammalian cells. Mutant C proteins unable to bind AIP1/Alix failed to accelerate the release of virus-like particles from cells. Furthermore, overexpression of AIP1/Alix enhanced SeV budding from infected cells in a C-protein-dependent manner, while the release of nucleocapsid-free empty virions was also enhanced. Finally, AIP1/Alix depletion by small interfering RNA resulted in suppression of SeV budding. The results of this study suggest that AIP1/Alix plays a role in efficient SeV budding and that the SeV C protein facilitates virus budding through interaction with AIP1/Alix.

Ultrastructural Alterations in Hela Cells Induced by Spider Venom

1967 ◽  
Vol 25 ◽  
pp. 20-21
Author(s):  
Dale E. McClendon ◽  
Paul N. Morgan ◽  
Bernard L. Soloff
Keyword(s):  
Growth Medium ◽  
Mammalian Cells ◽  
Carcinoma Cells ◽  
Venom Protein ◽  
Sterile Saline ◽  
Carbon Coated ◽  
Available Information ◽  
Loxosceles Reclusa ◽  
Essential Growth ◽  
Solution Cultures

It has been observed that minute amounts of venom from the brown recluse spider, Loxosceles reclusa, are capable of producing cytotoxic changes in cultures of certain mammalian cells (Morgan and Felton, 1965). Since there is little available information concerning the effect of venoms on susceptible cells, we have attempted to characterize, at the electron microscope level, the cytotoxic changes produced by the venom of this spider.Cultures of human epithelial carcinoma cells, strain HeLa, were initiated on sterile, carbon coated coverslips contained in Leighton tubes. Each culture was seeded with approximately 1x105 cells contained in 1.5 ml of a modified Eagle's minimum essential growth medium prepared in Hank's balanced salt solution. Cultures were incubated at 36° C. for three days prior to the addition of venom. The venom was collected from female brown recluse spiders and diluted in sterile saline. Protein determinations on the venom-were made according to the spectrophotometric method of Waddell (1956). Approximately 10 μg venom protein per ml of fresh medium was added to each culture after discarding the old growth medium. Control cultures were treated similarly, except that no venom was added. All cultures were reincubated at 36° C.

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