scholarly journals A Conformational Transition Observed in Single HIV-1 Gag Molecules during In Vitro Assembly of Virus-Like Particles

2014 ◽  
Vol 88 (6) ◽  
pp. 3577-3585 ◽  
Author(s):  
J. B. Munro ◽  
A. Nath ◽  
M. Farber ◽  
S. A. K. Datta ◽  
A. Rein ◽  
...  
Keyword(s):  
Conformational Transition ◽  
Virus Like Particles ◽  
In Vitro Assembly ◽  
Hiv 1

scholarly journals Effect of Small Polyanions on In Vitro Assembly of Selected Members of Alpha-, Beta- and Gammaretroviruses

Viruses ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 129
Author(s):  
Alžběta Dostálková ◽  
Barbora Vokatá ◽  
Filip Kaufman ◽  
Pavel Ulbrich ◽  
Tomáš Ruml ◽  
...  
Keyword(s):  
Equine Infectious Anemia Virus ◽  
Electron Tomography ◽  
Leukemia Virus ◽  
Virus Like Particles ◽  
Rous Sarcoma ◽  
Sarcoma Virus ◽  
Anemia Virus ◽  
Immature Virus ◽  
Hiv 1

The assembly of a hexameric lattice of retroviral immature particles requires the involvement of cell factors such as proteins and small molecules. A small, negatively charged polyanionic molecule, myo-inositol hexaphosphate (IP6), was identified to stimulate the assembly of immature particles of HIV-1 and other lentiviruses. Interestingly, cryo-electron tomography analysis of the immature particles of two lentiviruses, HIV-1 and equine infectious anemia virus (EIAV), revealed that the IP6 binding site is similar. Based on this amino acid conservation of the IP6 interacting site, it is presumed that the assembly of immature particles of all lentiviruses is stimulated by IP6. Although this specific region for IP6 binding may be unique for lentiviruses, it is plausible that other retroviral species also recruit some small polyanion to facilitate the assembly of their immature particles. To study whether the assembly of retroviruses other than lentiviruses can be stimulated by polyanionic molecules, we measured the effect of various polyanions on the assembly of immature virus-like particles of Rous sarcoma virus (RSV), a member of alpharetroviruses, Mason-Pfizer monkey virus (M-PMV) representative of betaretroviruses, and murine leukemia virus (MLV), a member of gammaretroviruses. RSV, M-PMV and MLV immature virus-like particles were assembled in vitro from truncated Gag molecules and the effect of selected polyanions, myo-inostol hexaphosphate, myo-inositol, glucose-1,6-bisphosphate, myo-inositol hexasulphate, and mellitic acid, on the particles assembly was quantified. Our results suggest that the assembly of immature particles of RSV and MLV was indeed stimulated by the presence of myo-inostol hexaphosphate and myo-inositol, respectively. In contrast, no effect on the assembly of M-PMV as a betaretrovirus member was observed.

scholarly journals The Retroviral Capsid Domain Dictates Virion Size, Morphology, and Coassembly of Gag into Virus-Like Particles

2005 ◽  
Vol 79 (21) ◽  
pp. 13463-13472 ◽  
Author(s):  
Danso Ako-Adjei ◽  
Marc C. Johnson ◽  
Volker M. Vogt
Keyword(s):  
Nuclear Export ◽  
Leukemia Virus ◽  
Particle Diameter ◽  
Particle Morphology ◽  
Structural Protein ◽  
Wild Type ◽  
Virus Like Particles ◽  
Gag Proteins ◽  
Hiv 1

ABSTRACT The retroviral structural protein, Gag, is capable of independently assembling into virus-like particles (VLPs) in living cells and in vitro. Immature VLPs of human immunodeficiency virus type 1 (HIV-1) and of Rous sarcoma virus (RSV) are morphologically distinct when viewed by transmission electron microscopy (TEM). To better understand the nature of the Gag-Gag interactions leading to these distinctions, we constructed vectors encoding several RSV/HIV-1 chimeric Gag proteins for expression in either insect cells or vertebrate cells. We used TEM, confocal fluorescence microscopy, and a novel correlative scanning EM (SEM)-confocal microscopy technique to study the assembly properties of these proteins. Most chimeric proteins assembled into regular VLPs, with the capsid (CA) domain being the primary determinant of overall particle diameter and morphology. The presence of domains between matrix and CA also influenced particle morphology by increasing the spacing between the inner electron-dense ring and the VLP membrane. Fluorescently tagged versions of wild-type RSV, HIV-1, or murine leukemia virus Gag did not colocalize in cells. However, wild-type Gag proteins colocalized extensively with chimeric Gag proteins bearing the same CA domain, implying that Gag interactions are mediated by CA. A dramatic example of this phenomenon was provided by a nuclear export-deficient chimera of RSV Gag carrying the HIV-1 CA domain, which by itself localized to the nucleus but relocalized to the cytoplasm in the presence of wild type HIV-1 Gag. Wild-type and chimeric Gag proteins were capable of coassembly into a single VLP as viewed by correlative fluorescence SEM if, and only if, the CA domain was derived from the same virus. These results imply that the primary selectivity of Gag-Gag interactions is determined by the CA domain.

scholarly journals A Novel MVA-Based HIV Vaccine Candidate (MVA-gp145-GPN) Co-Expressing Clade C Membrane-Bound Trimeric gp145 Env and Gag-Induced Virus-Like Particles (VLPs) Triggered Broad and Multifunctional HIV-1-Specific T Cell and Antibody Responses

Viruses ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 160 ◽  
Author(s):  
Beatriz Perdiguero ◽  
Cristina Sánchez-Corzo ◽  
Carlos Sorzano ◽  
Lidia Saiz ◽  
Pilar Mediavilla ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Vaccine Candidate ◽  
Hiv Vaccine ◽  
Replication Capacity ◽  
Virus Like Particles ◽  
Modified Vaccinia Virus Ankara ◽  
Clade C ◽  
Membrane Bound ◽  
Hiv 1

The development of an effective Human Immunodeficiency Virus (HIV) vaccine that is able to stimulate both the humoral and cellular HIV-1-specific immune responses remains a major priority challenge. In this study, we described the generation and preclinical evaluation of single and double modified vaccinia virus Ankara (MVA)-based candidates expressing the HIV-1 clade C membrane-bound gp145(ZM96) trimeric protein and/or the Gag(ZM96)-Pol-Nef(CN54) (GPN) polyprotein that was processed to form Gag-induced virus-like particles (VLPs). In vitro characterization of MVA recombinants revealed the stable integration of HIV-1 genes without affecting its replication capacity. In cells that were infected with Env-expressing viruses, the gp145 protein was inserted into the plasma membrane exposing critical epitopes that were recognized by broadly neutralizing antibodies (bNAbs), whereas Gag-induced VLPs were released from cells that were infected with GPN-expressing viruses. VLP particles as well as purified MVA virions contain Env and Gag visualized by immunoelectron microscopy and western-blot of fractions that were obtained after detergent treatments of purified virus particles. In BALB/c mice, homologous MVA-gp145-GPN prime/boost regimen induced broad and polyfunctional Env- and Gag-specific CD4 T cells and antigen-specific T follicular helper (Tfh) and Germinal Center (GC) B cells, which correlated with robust HIV-1-specific humoral responses. Overall, these results support the consideration of MVA-gp145-GPN vector as a potential vaccine candidate against HIV-1.

A Quantitative Description of In Vitro Assembly of Human Papillomavirus 16 Virus-Like Particles

2008 ◽  
Vol 381 (1) ◽  
pp. 229-237 ◽  
Author(s):  
Santanu Mukherjee ◽  
Marc V. Thorsteinsson ◽  
Lauren Beth Johnston ◽  
Pete A. DePhillips ◽  
Adam Zlotnick
Keyword(s):  
Human Papillomavirus ◽  
Quantitative Description ◽  
Virus Like Particles ◽  
Human Papillomavirus 16 ◽  
In Vitro Assembly

scholarly journals Electrostatic repulsion between HIV-1 capsid proteins modulates hexamer plasticity and in vitro assembly

2010 ◽  
pp. NA-NA
Author(s):  
Sonia Brun ◽  
Laurent Chaloin ◽  
Bernard Gay ◽  
Eric Bernard ◽  
Christian Devaux ◽  
...  
Keyword(s):  
Capsid Proteins ◽  
Electrostatic Repulsion ◽  
In Vitro Assembly ◽  
Hiv 1

scholarly journals Envelope Lipids Regulate the In Vitro Assembly of the HIV-1 Capsid

2008 ◽  
Vol 94 (2) ◽  
pp. L08-L10 ◽  
Author(s):  
Francisco N. Barrera ◽  
Marta del Álamo ◽  
Mauricio G. Mateu ◽  
José L. Neira
Keyword(s):  
In Vitro Assembly ◽  
Hiv 1

Characterization of a reactive groove in HIV-1 capsid protein defined by an in vitro assembly inhibitor

2007 ◽  
Vol 2007 (Spring) ◽  
Author(s):  
Vanda Bartonova ◽  
Jana Sticht ◽  
Peter Sehr ◽  
Joe Lewis ◽  
Hans-Georg Kraeusslich
Keyword(s):  
Capsid Protein ◽  
In Vitro Assembly ◽  
Hiv 1

scholarly journals In vitro assembly of polymorphic virus-like particles from the capsid protein of a nodavirus

Virology ◽  
2016 ◽  
Vol 496 ◽  
pp. 106-115 ◽  
Author(s):  
Saumya Bajaj ◽  
Manidipa Banerjee
Keyword(s):  
Capsid Protein ◽  
Virus Like Particles ◽  
In Vitro Assembly
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