scholarly journals Phototracking Vaccinia Virus Transport Reveals Dynamics of Cytoplasmic Dispersal and a Requirement for A36R and F12L for Exit from the Site of Wrapping

Viruses ◽  
2018 ◽  
Vol 10 (8) ◽  
pp. 390
Author(s):  
Helena Lynn ◽  
Liam Howell ◽  
Russell Diefenbach ◽  
Timothy Newsome
Keyword(s):  
Vaccinia Virus ◽  
Viral Infections ◽  
Cell Periphery ◽  
Virus Transport ◽  
Anterograde Transport ◽  
Recombinant Viruses ◽  
Virus Particles ◽  
Viral Particles ◽  
Viral Morphogenesis ◽  
Golgi Network

The microtubule cytoskeleton is a primary organizer of viral infections for delivering virus particles to their sites of replication, establishing and maintaining subcellular compartments where distinct steps of viral morphogenesis take place, and ultimately dispersing viral progeny. One of the best characterized examples of virus motility is the anterograde transport of the wrapped virus form of vaccinia virus (VACV) from the trans-Golgi network (TGN) to the cell periphery by kinesin-1. Yet many aspects of this transport event are elusive due to the speed of motility and the challenges of imaging this stage at high resolution over extended time periods. We have established a novel imaging technology to track virus transport that uses photoconvertible fluorescent recombinant viruses to track subsets of virus particles from their site of origin and determine their destination. Here we image virus exit from the TGN and their rate of egress to the cell periphery. We demonstrate a role for kinesin-1 engagement in regulating virus exit from the TGN by removing A36 and F12 function, critical viral mediators of kinesin-1 recruitment to virus particles. Phototracking viral particles and components during infection is a powerful new imaging approach to elucidate mechanisms of virus replication.

scholarly journals An investigation of incorporation of cellular antigens into vaccinia virus particles

2002 ◽  
Vol 83 (10) ◽  
pp. 2347-2359 ◽  
Author(s):  
Oliver Krauss ◽  
Ruth Hollinshead ◽  
Michael Hollinshead ◽  
Geoffrey L. Smith
Keyword(s):  
Vaccinia Virus ◽  
Specific Cell ◽  
Virus Particles ◽  
Enveloped Virus ◽  
Membrane Compartments ◽  
Specific Proteins ◽  
Intermediate Compartment ◽  
Low Levels ◽  
Golgi Network ◽  
Exocytic Pathway

Vaccinia virus (VV) infection produces several types of virus particle called intracellular mature virus (IMV), intracellular enveloped virus (IEV), cell-associated enveloped virus (CEV) and extracellular enveloped virus (EEV). Some cellular antigens are associated with EEV and these vary with the cell type used to grow the virus. To investigate if specific cell antigens are associated with VV particles, and to address the origin of membranes used to envelope IMV and IEV/CEV/EEV, we have studied whether cell antigens and foreign antigens expressed by recombinant VVs are incorporated into VV particles. Membrane proteins that are incorporated into the endoplasmic reticulum (ER), intermediate compartment (IC), cis/medial-Golgi, trans-Golgi network (TGN) or plasma membrane were not detected in purified IMV particles. In contrast, proteins present in the TGN or membrane compartments further downstream in the exocytic pathway co-purify with EEV particles when analysed by immunoblotting. Immunoelectron microscopy found only low levels of these proteins in IEV, CEV/EEV. The incorporation of foreign antigens into VV particles was not affected by loss of individual IEV or EEV-specific proteins or by redirection of B5R to the ER. These data suggest that (i) host cell antigens are excluded from the lipid envelope surrounding the IMV particle and (ii) membranes of the ER, IC and cis/medial-Golgi are not used to wrap IMV particles to form IEV. Lastly, the VV haemagglutinin was absent from one-third of IEV and CEV/EEV particles, whereas other EEV antigens were present in all these virions.

scholarly journals Movements of vaccinia virus intracellular enveloped virions with GFP tagged to the F13L envelope protein

2001 ◽  
Vol 82 (11) ◽  
pp. 2747-2760 ◽  
Author(s):  
María M. Geada ◽  
Inmaculada Galindo ◽  
María M. Lorenzo ◽  
Beatriz Perdiguero ◽  
Rafael Blasco
Keyword(s):  
Cell Surface ◽  
Vaccinia Virus ◽  
Intracellular Transport ◽  
Virus Transport ◽  
Enveloped Virus ◽  
Vaccinia Viruses ◽  
Butanedione Monoxime ◽  
Golgi Network ◽  
Dependent Mechanism

Vaccinia virus produces several forms of infectious virions. Intracellular mature virions (IMV) assemble in areas close to the cell nucleus. Some IMV acquire an envelope from intracellular membranes derived from the trans-Golgi network, producing enveloped forms found in the cytosol (intracellular enveloped virus; IEV), on the cell surface (cell-associated enveloped virus) or free in the medium (extracellular enveloped virus; EEV). Blockage of IMV envelopment inhibits transport of virions to the cell surface, indicating that enveloped virus forms are required for virion movement from the Golgi area. To date, the induction of actin tails that propel IEV is the only well-characterized mechanism for enveloped virus transport. However, enveloped virus transport and release occur under conditions where actin tails are not formed. In order to study these events, recombinant vaccinia viruses were constructed with GFP fused to the most abundant protein in the EEV envelope, P37 (F13L). The P37–GFP fusion, like normal P37, accumulated in the Golgi area and was incorporated efficiently into enveloped virions. These recombinants allowed the monitoring of enveloped virus movements in vivo. In addition to a variety of relatively slow movements (<0·4 μm/s), faster, saltatory movements both towards and away from the Golgi area were observed. These movements were different from those dependent on actin tails and were inhibited by the microtubule-disrupting drug nocodazole, but not by the myosin inhibitor 2,3-butanedione monoxime. Video microscopy (5 frames per s) revealed that saltatory movements had speeds of up to, and occasionally more than, 3 μm/s. These results suggest that a second, microtubule-dependent mechanism exists for intracellular transport of enveloped vaccinia virions.

scholarly journals Directional Spread of Surface-Associated Retroviruses Regulated by Differential Virus-Cell Interactions

2010 ◽  
Vol 84 (7) ◽  
pp. 3248-3258 ◽  
Author(s):  
Nathan M. Sherer ◽  
Jing Jin ◽  
Walther Mothes
Keyword(s):  
Cell Surface ◽  
Viral Infections ◽  
Leukemia Virus ◽  
Cell Interactions ◽  
Target Cells ◽  
Specific Cell ◽  
Cell Periphery ◽  
Surface Receptors ◽  
Virus Particles ◽  
Infected Cells

ABSTRACT The spread of viral infections involves the directional progression of virus particles from infected cells to uninfected target cells. Prior to entry, the binding of virus particles to specific cell surface receptors can trigger virus surfing, an actin-dependent lateral transport of viruses toward the cell body (M. J. Lehmann et al., J. Cell Biol. 170:317-325, 2005; M. Schelhaas, et al., PLoS Pathog. 4:e1000148, 2008; J. L. Smith, D. S. Lidke, and M. A. Ozbun, Virology 381:16-21, 2008). Here, we have used live-cell imaging to demonstrate that for cells chronically infected with the gammaretrovirus murine leukemia virus in which receptor has been downregulated, a significant portion of completely assembled virus particles are not immediately released into the supernatant but retain long-term association with the cell surface. Retention can be attributed, at least in part, to nonspecific particle attachment to cell surface glycosylaminoglycans. In contrast to virus surfing, viruses retained at the surface of infected cells undergo a lateral motility that is random and actin independent. This diffusive motility can be abruptly halted and converted into inward surfing after treatment with Polybrene, a soluble cation that increases virus-cell adsorption. In the absence of Polybrene, particle diffusion allows for an outward flow of viruses to the infected cell periphery. Peripheral particles are readily captured by and transmitted to neighboring uninfected target cells in a directional fashion. These data demonstrate a surface-based mechanism for the directional spread of viruses regulated by differential virus-cell interactions.

scholarly journals Bi-reporter vaccinia virus for tracking viral infections in vitro and in vivo

2021 ◽  
Author(s):  
Kevin Chiem ◽  
María Lorenzo ◽  
Javier Rangel-Moreno ◽  
Maria de la Luz Garcia-Hernandez ◽  
Jun-Gyu Park ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Viral Infections ◽  
Viral Vector ◽  
Ex Vivo ◽  
Reporter Genes ◽  
Luciferase Reporter ◽  
Recombinant Viruses ◽  
Antibody Staining

Recombinant viruses expressing reporter genes allow visualization and quantification of viral infections and can be used as valid surrogates to identify the presence of the virus in infected cells and animal models. However, one of the limitations of recombinant viruses expressing reporter genes is the use of either fluorescent or luciferase proteins that are used alternatively for different purposes. Vaccinia virus (VV) is widely used as a viral vector, including recombinant (r)VV singly expressing either fluorescent or luciferase reporter genes that are useful for specific purposes. In this report, we engineered two novel rVV stably expressing both fluorescent (Scarlet or GFP) and luciferase (Nluc) reporter genes from different loci in the viral genome. In vitro, these bi-reporter expressing rVV have similar growth kinetics and plaque phenotype than those of the parental WR VV isolate. In vivo, rVV Nluc/Scarlet and rVV Nluc/GFP effectively infected mice and were easily detected using in vivo imaging systems (IVIS) and ex vivo in the lungs from infected mice. We used these bi-reporter expressing rVV to assess viral pathogenesis, infiltration of immune cells in the lungs, and to directly identify the different subsets of cells infected by VV in the absence of antibody staining. Collectively, these rVV expressing two reporter genes open the feasibility to study the biology of viral infections in vitro and in vivo, including host-pathogen interactions and dynamics or tropism of viral infections. Moreover, they represent an excellent approach for the discovery of new prophylactics and/or therapeutics for the treatment of poxvirus infections.

scholarly journals Protease Inhibitors as Antiviral Agents

1998 ◽  
Vol 11 (4) ◽  
pp. 614-627 ◽  
Author(s):  
A. K. Patick ◽  
K. E. Potts
Keyword(s):  
Protease Inhibitors ◽  
Viral Infections ◽  
Critical Role ◽  
Antiviral Agents ◽  
Antiviral Agent ◽  
Structural Proteins ◽  
Viral Protease ◽  
Virus Particles ◽  
Viral Proteases ◽  
Viral Polyprotein

SUMMARY Currently, there are a number of approved antiviral agents for use in the treatment of viral infections. However, many instances exist in which the use of a second antiviral agent would be beneficial because it would allow the option of either an alternative or a combination therapeutic approach. Accordingly, virus-encoded proteases have emerged as new targets for antiviral intervention. Molecular studies have indicated that viral proteases play a critical role in the life cycle of many viruses by effecting the cleavage of high-molecular-weight viral polyprotein precursors to yield functional products or by catalyzing the processing of the structural proteins necessary for assembly and morphogenesis of virus particles. This review summarizes some of the important general features of virus-encoded proteases and highlights new advances and/or specific challenges that are associated with the research and development of viral protease inhibitors. Specifically, the viral proteases encoded by the herpesvirus, retrovirus, hepatitis C virus, and human rhinovirus families are discussed.

scholarly journals Targeting Viral Surface Proteins through Structure-Based Design

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1320
Author(s):  
Yogesh B Narkhede ◽  
Karen J Gonzalez ◽  
Eva-Maria Strauch
Keyword(s):  
Public Health ◽  
Viral Infections ◽  
Respiratory Viruses ◽  
Surface Proteins ◽  
Host Cells ◽  
Viral Fusion ◽  
Health It ◽  
Viral Particles ◽  
Pathogenic Viruses ◽  
Viral Surface

The emergence of novel viral infections of zoonotic origin and mutations of existing human pathogenic viruses represent a serious concern for public health. It warrants the establishment of better interventions and protective therapies to combat the virus and prevent its spread. Surface glycoproteins catalyzing the fusion of viral particles and host cells have proven to be an excellent target for antivirals as well as vaccines. This review focuses on recent advances for computational structure-based design of antivirals and vaccines targeting viral fusion machinery to control seasonal and emerging respiratory viruses.

scholarly journals Influence of Meteorological Conditions and Aerosol Properties on the COVID-19 Contamination of the Population in Coastal and Continental Areas in France: Study of Offshore and Onshore Winds

Atmosphere ◽  
2021 ◽  
Vol 12 (4) ◽  
pp. 523
Author(s):  
Jacques Piazzola ◽  
William Bruch ◽  
Christelle Desnues ◽  
Philippe Parent ◽  
Christophe Yohia ◽  
...  
Keyword(s):  
Atmospheric Transport ◽  
Weather Conditions ◽  
Meteorological Conditions ◽  
Sea Spray ◽  
Atmospheric Conditions ◽  
Virus Transport ◽  
Virus Propagation ◽  
Viral Particles ◽  
The Social ◽  
Aerosol Properties

Human behaviors probably represent the most important causes of the SARS-Cov-2 virus propagation. However, the role of virus transport by aerosols—and therefore the influence of atmospheric conditions (temperature, humidity, type and concentration of aerosols)—on the spread of the epidemic remains an open and still debated question. This work aims to study whether or not the meteorological conditions related to the different aerosol properties in continental and coastal urbanized areas might influence the atmospheric transport of the SARS-Cov-2 virus. Our analysis focuses on the lockdown period to reduce the differences in the social behavior and highlight those of the weather conditions. As an example, we investigated the contamination cases during March 2020 in two specific French areas located in both continental and coastal areas with regard to the meteorological conditions and the corresponding aerosol properties, the optical depth (AOD) and the Angstrom exponent provided by the AERONET network. The results show that the analysis of aerosol ground-based data can be of interest to assess a virus survey. We found that moderate to strong onshore winds occurring in coastal regions and inducing humid environment and large sea-spray production episodes coincides with smaller COVID-19 contamination rates. We assume that the coagulation of SARS-Cov-2 viral particles with hygroscopic salty sea-spray aerosols might tend to inhibit its viral infectivity via possible reaction with NaCl, especially in high relative humidity environments typical of maritime sites.

Effect of detergents and chemicals on purified vaccinia virus: analysis by SDS polyacrylamide gel electrophoresis and electron microscopy

1977 ◽  
Vol 23 (3) ◽  
pp. 240-252 ◽  
Author(s):  
J. Boisvert ◽  
T. Yamamoto
Keyword(s):  
Electron Microscopy ◽  
Vaccinia Virus ◽  
Gel Electrophoresis ◽  
Alkali Treatment ◽  
Guanidine Hydrochloride ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Ammonium Bromide ◽  
Molecular Weights ◽  
Viral Particles

Vaccinia virus particles were dissociated into their constituent polypeptides and analysed by sodium dodecyl sulfate (SDS) gel electrophoresis. Thirty-three distinct polypeptide bands were identified and their molecular weights ranged between 11 000 and 150 000 daltons.Specific staining of gels containing polypeptides of dissociated virions revealed the presence of eight glycopeptides. No lipopeptides were detected.Analysis of chemical extracts (urea, guanidine hydrochloride, and alkali treatment) of the virus by SDS gel electrophoresis indicated that a total of 10 to 14 different polypeptides ranging in molecular weights from 11 000 to 70 000 daltons were solubilized.Analysis of detergent extracts and of the remains of extracted viral particles has shown that the detergent Nonidet P-40 (NP-40) solubilized a total of 11 polypeptides of which 6 were glycopeptides. The other detergents sodium deoxycholate (SDC) and cetyl trimethyl ammonium bromide (CTAB) were not as selective, both solubilizing more than 25 of the polypeptides composing the virus. Gel electrophoresis results also indicated that most of the small molecular weight (11 000–70 000 daltons) polypeptides were readily solubilized by NP-40, SDC, and CTAB, while those with molecular weights of 70 000 daltons and higher were not well solubilized.The effects of detergents were also analysed by electron microscopy. Evidence was obtained for subpopulations of viral particles having different susceptibility to detergent extraction.

scholarly journals Identification and Characterization of a Poliovirus Capsid Mutant with Enhanced Thermal Stability

2018 ◽  
Vol 93 (6) ◽  
Author(s):  
Y Nguyen ◽  
Palmy R. Jesudhasan ◽  
Elizabeth R. Aguilera ◽  
Julie K. Pfeiffer
Keyword(s):  
Thermal Stability ◽  
High Temperatures ◽  
Oral Route ◽  
Mutant Virus ◽  
Single Amino Acid ◽  
Bacterial Surface ◽  
Virus Particles ◽  
Heat Resistant ◽  
Viral Particles ◽  
Very High

ABSTRACTEnteric viruses, including poliovirus, are spread by the fecal-oral route. In order to persist and transmit to a new host, enteric virus particles must remain stable once they are in the environment. Environmental stressors such as heat and disinfectants can inactivate virus particles and prevent viral transmission. It has been previously demonstrated that bacteria or bacterial surface glycans can enhance poliovirus virion stability and limit inactivation from heat or bleach. While investigating the mechanisms underlying bacterially enhanced virion thermal stability, we identified and characterized a poliovirus (PV) mutant with increased resistance to heat inactivation. The M132V mutant harbors a single amino acid change in the VP1 capsid coding that is sufficient to confer heat resistance but not bleach resistance. Although the M132V virus was stable in the absence of bacteria or feces at most temperatures, M132V virus was stabilized by feces at very high temperatures. M132V PV had reduced specific infectivity and RNA uncoating compared with those of wild-type (WT) PV, but viral yields in HeLa cells were similar. In orally inoculated mice, M132V had a slight fitness cost since fecal titers were lower and 12.5% of fecal viruses reverted to the WT. Overall, this work sheds light on factors that influence virion stability and fitness.IMPORTANCEViruses spread by the fecal-oral route need to maintain viability in the environment to ensure transmission. Previous work indicated that bacteria and bacterial surface polysaccharides can stabilize viral particles and enhance transmission. To explore factors that influence viral particle stability, we isolated a mutant poliovirus that is heat resistant. This mutant virus does not require feces for stability at most temperatures but can be stabilized by feces at very high temperatures. Even though the mutant virus is heat resistant, it is susceptible to inactivation by treatment with bleach. This work provides insight into how viral particles maintain infectivity in the environment.

scholarly journals Abundant Defective Viral Particles Budding from Microglia in the Course of Retroviral Spongiform Encephalopathy

2000 ◽  
Vol 74 (4) ◽  
pp. 1775-1780 ◽  
Author(s):  
Regine Hansen ◽  
Stefanie Czub ◽  
Evi Werder ◽  
Jens Herold ◽  
Georg Gosztonyi ◽  
...  
Keyword(s):  
Infectious Virus ◽  
Protein Translocation ◽  
Neuronal Damage ◽  
Ex Vivo ◽  
Spongiform Encephalopathy ◽  
Microglia Cell ◽  
Virus Particles ◽  
Viral Particles ◽  
Type C ◽  
Retroviral Infections

ABSTRACT A pathogenetic hallmark of retroviral neurodegeneration is the affinity of neurovirulent retroviruses for microglia cells, while degenerating neurons are excluded from retroviral infections. Microglia isolated ex vivo from rats peripherally infected with a neurovirulent retrovirus released abundant mature type C virions; however, infectivity associated with microglia was very low. In microglia, viral transcription was unaffected but envelope proteins were insufficiently cleaved into mature viral proteins and were not detected on the microglia cell surface. These microglia-specific defects in envelope protein translocation and processing not only may have prevented formation of infectious virus particles but also may have caused further cellular defects in microglia with the consequence of indirect neuronal damage. It is conceivable that similar events play a role in neuro-AIDS.

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