scholarly journals Cylindrical inclusion protein of potato virus A is associated with a subpopulation of particles isolated from infected plants

2008 ◽  
Vol 89 (3) ◽  
pp. 829-838 ◽  
Author(s):  
Rasa Gabrenaite-Verkhovskaya ◽  
Igor A. Andreev ◽  
Natalia O. Kalinina ◽  
Lesley Torrance ◽  
Michael E. Taliansky ◽  
...  
Keyword(s):  
Potato Virus ◽  
Immunogold Labelling ◽  
Cylindrical Inclusion ◽  
Force Microscopy ◽  
Virus Particles ◽  
Potato Virus A ◽  
Atomic Force ◽  
Inclusion Protein ◽  
Cylindrical Inclusion Protein ◽  
Gradient Separation

Potato virus A (PVA) particles were purified by centrifugation through a 30 % sucrose cushion and the pellet (P1) was resuspended and sedimented through a 5–40 % sucrose gradient. The gradient separation resulted in two different virus particle populations: a virus fraction (F) that formed a band in the gradient and one that formed a pellet (P2) at the bottom of the gradient. All three preparations contained infectious particles that retained their integrity when visualized by electron microscopy (EM). Western blotting of the P1 particles revealed that the viral RNA helicase, cylindrical inclusion protein (CI), co-purified with virus particles. This result was confirmed with co-immunoprecipitation experiments. CI was detected in P2 particle preparations, whereas F particles were devoid of detectable amounts of CI. ATPase activity was detected in all three preparations with the greatest amount in P2. Results from immunogold-labelling EM experiments suggested that a fraction of the CI present in the preparations was localized to one end of the virion. Atomic force microscopy (AFM) studies showed that P1 and P2 contained intact particles, some of which had a protruding tip structure at one end, whilst F virions were less stable and mostly appeared as beaded structures under the conditions of AFM. The RNA of the particles in F was translated five to ten times more efficiently than RNA from P2 particles when these preparations were subjected to translation in wheat-germ extracts. The results are discussed in the context of a model for CI-mediated functions.

Reactivity of Two Monoclonal Antibodies to the Cylindrical Inclusion Protein of Potato Virus Y

1994 ◽  
Vol 141 (2) ◽  
pp. 186-194
Author(s):  
G. Boudazin ◽  
C. Vergnet ◽  
B. Gélie ◽  
M. Meyer ◽  
J. Grosclaude ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Cylindrical Inclusion ◽  
Inclusion Protein ◽  
Cylindrical Inclusion Protein

Atomic force microscopy of potato virus A

2008 ◽  
Vol 70 (2) ◽  
pp. 199-201 ◽  
Author(s):  
E. A. Obraztsova ◽  
N. O. Kalinina ◽  
M. E. Taliansky ◽  
R. Gabrenaite-Verkhovskaya ◽  
K. Makinen ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Potato Virus ◽  
Force Microscopy ◽  
Potato Virus A ◽  
Atomic Force

scholarly journals Atomic Force Microscopy Investigation of Human Immunodeficiency Virus (HIV) and HIV-Infected Lymphocytes

2003 ◽  
Vol 77 (22) ◽  
pp. 11896-11909 ◽  
Author(s):  
Y. G. Kuznetsov ◽  
J. G. Victoria ◽  
W. E. Robinson ◽  
A. McPherson
Keyword(s):  
Human Immunodeficiency Virus ◽  
Atomic Force Microscopy ◽  
Nucleic Acid ◽  
Matrix Protein ◽  
Human Lymphocytes ◽  
Tween 20 ◽  
Force Microscopy ◽  
Virus Particles ◽  
Atomic Force ◽  
Immunodeficiency Virus

ABSTRACT Isolated human immunodeficiency virus (HIV) and HIV-infected human lymphocytes in culture have been imaged for the first time by atomic force microscopy (AFM). Purified virus particles spread on glass substrates are roughly spherical, reasonably uniform, though pleomorphic in appearance, and have diameters of about 120 nm. Similar particles are also seen on infected cell surfaces, but morphologies and sizes are considerably more varied, possibly a reflection of the budding process. The surfaces of HIV particles exhibit “tufts” of protein, presumably gp120, which do not physically resemble spikes. The protein tufts, which number about 100 per particle, have average diameters of about 200 Å, but with a large variance. They likely consist of arbitrary associations of small numbers of gp120 monomers on the surface. In examining several hundred virus particles, we found no evidence that the gp120 monomers form threefold symmetric trimers. Although >95% of HIV-infected H9 lymphocytic cells were producing HIV antigens by immunofluorescent assay, most lymphocytes displayed few or no virus on their surfaces, while others were almost covered by a hundred or more viruses, suggesting a dependence on cell cycle or physiology. HIV-infected cells treated with a viral protease inhibitor and their progeny viruses were also imaged by AFM and were indistinguishable from untreated virions. Isolated HIV virions were disrupted by exposure to mild neutral detergents (Tween 20 and CHAPS) at concentrations from 0.25 to 2.0%. Among the products observed were intact virions, the remnants of completely degraded virions, and partially disrupted particles that lacked sectors of surface proteins as well as virions that were split or broken open to reveal their empty interiors. Capsids containing nucleic acid were not seen, suggesting that the capsids were even more fragile than the envelope and were totally degraded and lost. From these images, a good estimate of the thickness of the envelope protein-membrane-matrix protein outer shell of the virion was obtained. Treatment with even low concentrations (<0.1%) of sodium dodecyl sulfate completely destroyed all virions but produced many interesting products, including aggregates of viral proteins with strands of nucleic acid.

scholarly journals Nanoscale Wetting of Single Viruses

Molecules ◽  
2021 ◽  
Vol 26 (17) ◽  
pp. 5184
Author(s):  
Annalisa Calò ◽  
Aitziber Eleta-Lopez ◽  
Thierry Ondarçuhu ◽  
Albert Verdaguer ◽  
Alexander M. Bittner
Keyword(s):  
Relative Humidity ◽  
Viral Infections ◽  
High Sensitivity ◽  
Water Layer ◽  
Dynamic Force ◽  
Force Microscopy ◽  
Virus Particles ◽  
Ambient Relative Humidity ◽  
Atomic Force ◽  
Condensed Water

The epidemic spread of many viral infections is mediated by the environmental conditions and influenced by the ambient humidity. Single virus particles have been mainly visualized by atomic force microscopy (AFM) in liquid conditions, where the effect of the relative humidity on virus topography and surface cannot be systematically assessed. In this work, we employed multi-frequency AFM, simultaneously with standard topography imaging, to study the nanoscale wetting of individual Tobacco Mosaic virions (TMV) from ambient relative humidity to water condensation (RH > 100%). We recorded amplitude and phase vs. distance curves (APD curves) on top of single virions at various RH and converted them into force vs. distance curves. The high sensitivity of multifrequency AFM to visualize condensed water and sub-micrometer droplets, filling gaps between individual TMV particles at RH > 100%, is demonstrated. Dynamic force spectroscopy allows detecting a thin water layer of thickness ⁓1 nm, adsorbed on the outer surface of single TMV particles at RH < 60%.

scholarly journals Laser spectroscopic technique for direct identification of a single virus I: FASTER CARS

2020 ◽  
Vol 117 (45) ◽  
pp. 27820-27824 ◽  
Author(s):  
Volker Deckert ◽  
Tanja Deckert-Gaudig ◽  
Dana Cialla-May ◽  
Jürgen Popp ◽  
Roland Zell ◽  
...  
Keyword(s):  
Clinical Sample ◽  
H1n1 Influenza ◽  
Spectroscopic Technique ◽  
Spectroscopic Techniques ◽  
Force Microscopy ◽  
Virus Particles ◽  
Detection Techniques ◽  
Atomic Force ◽  
Enhanced Resolution ◽  
Anti Stokes

From the famous 1918 H1N1 influenza to the present COVID-19 pandemic, the need for improved viral detection techniques is all too apparent. The aim of the present paper is to show that identification of individual virus particles in clinical sample materials quickly and reliably is near at hand. First of all, our team has developed techniques for identification of virions based on a modular atomic force microscopy (AFM). Furthermore, femtosecond adaptive spectroscopic techniques with enhanced resolution via coherent anti-Stokes Raman scattering (FASTER CARS) using tip-enhanced techniques markedly improves the sensitivity [M. O. Scully,et al.,Proc. Natl. Acad. Sci. U.S.A.99, 10994–11001 (2002)].

Interface between Adjacent UV-Treated or Detergent-Treated Vaccinia virus Particles

2005 ◽  
Vol 11 (S03) ◽  
pp. 86-89 ◽  
Author(s):  
A. V. Cardoso ◽  
G. S. Trindade
Keyword(s):  
Atomic Force Microscopy ◽  
Vaccinia Virus ◽  
Particle Surface ◽  
Surface Interactions ◽  
Surface Interaction ◽  
Force Microscopy ◽  
Virus Particles ◽  
Atomic Force ◽  
Analysis Of The Images ◽  
Viral Surface

While investigating isolated or agglomerates of treated Vaccinia virus intracellular mature (IMV) particles in atomic force microscopy (AFM) equipment we noticed that in some occasions the enveloped particles had been totally disrupted, with the interior being spread around. We have also observed in these samples what appear to be some rather intriguing viral surface interactions. Instead of showing a clear division between individual virions the particles seem to be continuous at the interfaces that show coalescence. In order to understand what was happening we focused our attention on the analysis of the images of the interface between virions particles, trying to find out what was the explanation for such type of particle surface interaction and in which conditions it would take place.

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