scholarly journals Author response: Efficient support of virus-like particle assembly by the HIV-1 packaging signal

2018 ◽  
Author(s):  
Mauricio Comas-Garcia ◽  
Tomas Kroupa ◽  
Siddhartha AK Datta ◽  
Demetria P Harvin ◽  
Wei-Shau Hu ◽  
...  
Keyword(s):  
Author Response ◽  
Packaging Signal ◽  
Particle Assembly ◽  
Virus Like Particle ◽  
Hiv 1

scholarly journals Efficient support of virus-like particle assembly by the HIV-1 packaging signal

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Mauricio Comas-Garcia ◽  
Tomas Kroupa ◽  
Siddhartha AK Datta ◽  
Demetria P Harvin ◽  
Wei-Shau Hu ◽  
...  
Keyword(s):  
Virus Assembly ◽  
Genomic Rna ◽  
Packaging Signal ◽  
Structural Element ◽  
Particle Assembly ◽  
Genomic Rnas ◽  
Gag Protein ◽  
Virus Particles ◽  
Virus Like Particle ◽  
Hiv 1

The principal structural component of a retrovirus particle is the Gag protein. Retroviral genomic RNAs contain a ‘packaging signal’ (‘Ψ') and are packaged in virus particles with very high selectivity. However, if no genomic RNA is present, Gag assembles into particles containing cellular mRNA molecules. The mechanism by which genomic RNA is normally selected during virus assembly is not understood. We previously reported (<xref ref-type="bibr" rid="bib9">Comas-Garcia et al., 2017</xref>) that at physiological ionic strength, recombinant HIV-1 Gag binds with similar affinities to RNAs with or without Ψ, and proposed that genomic RNA is selectively packaged because binding to Ψ initiates particle assembly more efficiently than other RNAs. We now present data directly supporting this hypothesis. We also show that one or more short stretches of unpaired G residues are important elements of Ψ; Ψ may not be localized to a single structural element, but is probably distributed over >100 bases.

scholarly journals Author response: Dissection of specific binding of HIV-1 Gag to the 'packaging signal' in viral RNA

2017 ◽  
Author(s):  
Mauricio Comas-Garcia ◽  
Siddhartha AK Datta ◽  
Laura Baker ◽  
Rajat Varma ◽  
Prabhakar R Gudla ◽  
...  
Keyword(s):  
Specific Binding ◽  
Viral Rna ◽  
Author Response ◽  
Packaging Signal ◽  
Hiv 1

scholarly journals HIV-1 matrix protein repositioning in nucleocapsid region fails to confer virus-like particle assembly

Virology ◽  
2008 ◽  
Vol 378 (1) ◽  
pp. 97-104 ◽  
Author(s):  
Ching-Yuan Chang ◽  
Yu-Fen Chang ◽  
Shiu-Mei Wang ◽  
Ying-Tzu Tseng ◽  
Kuo-Jung Huang ◽  
...  
Keyword(s):  
Matrix Protein ◽  
Particle Assembly ◽  
Virus Like Particle ◽  
Hiv 1

scholarly journals HIV-1 Gag protein with or without p6 specifically dimerizes on the viral RNA packaging signal

2020 ◽  
Vol 295 (42) ◽  
pp. 14391-14401 ◽  
Author(s):  
Samantha Sarni ◽  
Banhi Biswas ◽  
Shuohui Liu ◽  
Erik D. Olson ◽  
Jonathan P. Kitzrow ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Viral Particle ◽  
Rna Binding ◽  
Packaging Signal ◽  
Independent Manner ◽  
Particle Assembly ◽  
Gag Protein ◽  
Viral Particle Assembly ◽  
Hiv 1 ◽  
Psi Rna

The HIV-1 Gag protein is responsible for genomic RNA (gRNA) packaging and immature viral particle assembly. Although the presence of gRNA in virions is required for viral infectivity, in its absence, Gag can assemble around cellular RNAs and form particles resembling gRNA-containing particles. When gRNA is expressed, it is selectively packaged despite the presence of excess host RNA, but how it is selectively packaged is not understood. Specific recognition of a gRNA packaging signal (Psi) has been proposed to stimulate the efficient nucleation of viral assembly. However, the heterogeneity of Gag–RNA interactions renders capturing this transient nucleation complex using traditional structural biology approaches challenging. Here, we used native MS to investigate RNA binding of wild-type (WT) Gag and Gag lacking the p6 domain (GagΔp6). Both proteins bind to Psi RNA primarily as dimers, but to a control RNA primarily as monomers. The dimeric complexes on Psi RNA require an intact dimer interface within Gag. GagΔp6 binds to Psi RNA with high specificity in vitro and also selectively packages gRNA in particles produced in mammalian cells. These studies provide direct support for the idea that Gag binding to Psi specifically promotes nucleation of Gag–Gag interactions at the early stages of immature viral particle assembly in a p6-independent manner.

scholarly journals HIV-1 Gag protein with or without p6 specifically dimerizes on the viral RNA packaging signal

2020 ◽  
Author(s):  
Samantha Sarni ◽  
Banhi Biswas ◽  
Shuohui Liu ◽  
Erik D. Olson ◽  
Jonathan P. Kitzrow ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Viral Particle ◽  
Rna Binding ◽  
Packaging Signal ◽  
Independent Manner ◽  
Particle Assembly ◽  
Gag Protein ◽  
Viral Particle Assembly ◽  
Hiv 1 ◽  
Psi Rna

AbstractThe HIV-1 Gag protein is responsible for genomic RNA (gRNA) packaging and immature viral particle assembly. While the presence of gRNA in virions is required for viral infectivity, in its absence, Gag can assemble around cellular RNAs and form particles resembling gRNA-containing particles. When gRNA is expressed, it is selectively packaged despite the presence of excess host RNA, but how it is selectively packaged is not understood. Specific recognition of a gRNA packaging signal (Psi) has been proposed to stimulate the efficient nucleation of viral assembly. However, the heterogeneity of Gag-RNA interactions renders capturing this transient nucleation complex using traditional structural biology approaches challenging. Here, we used native mass spectrometry to investigate RNA binding of wild-type Gag and Gag lacking the p6 domain (GagΔp6). Both proteins bind to Psi RNA primarily as dimers, but to a control RNA primarily as monomers. The dimeric complexes on Psi RNA require an intact dimer interface within Gag. GagΔp6 binds to Psi RNA with high specificity in vitro and also selectively packages gRNA in particles produced in mammalian cells. These studies provide direct support for the idea that Gag binding to Psi specifically nucleates Gag-Gag interactions at the early stages of immature viral particle assembly in a p6-independent manner.

scholarly journals Analysis of Human Immunodeficiency Virus Type 1 Gag Dimerization-Induced Assembly

2005 ◽  
Vol 79 (23) ◽  
pp. 14498-14506 ◽  
Author(s):  
Ayna Alfadhli ◽  
Tenzin Choesang Dhenub ◽  
Amelia Still ◽  
Eric Barklis
Keyword(s):  
Human Immunodeficiency Virus ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Protein Assembly ◽  
Particle Assembly ◽  
Gag Protein ◽  
Immunodeficiency Virus ◽  
Hiv 1 ◽  
Terminal Domains

ABSTRACT The nucleocapsid (NC) domains of retrovirus precursor Gag (PrGag) proteins play an essential role in virus assembly. Evidence suggests that NC binding to viral RNA promotes dimerization of PrGag capsid (CA) domains, which triggers assembly of CA N-terminal domains (NTDs) into hexamer rings that are interconnected by CA C-terminal domains. To examine the influence of dimerization on human immunodeficiency virus type 1 (HIV-1) Gag protein assembly in vitro, we analyzed the assembly properties of Gag proteins in which NC domains were replaced with cysteine residues that could be linked via chemical treatment. In accordance with the model that Gag protein pairing triggers assembly, we found that cysteine cross-linking or oxidation reagents induced the assembly of virus-like particles. However, efficient assembly also was observed to be temperature dependent or required the tethering of NTDs. Our results suggest a multistep pathway for HIV-1 Gag protein assembly. In the first step, Gag protein pairing through NC-RNA interactions or C-terminal cysteine linkage fosters dimerization. Next, a conformational change converts assembly-restricted dimers or small oligomers into assembly-competent ones. At the final stage, final particle assembly occurs, possibly through a set of larger intermediates.

Structure and Stability of Wild-type and Mutant RNA Internal Loops from the SL-1 Domain of the HIV-1 Packaging Signal

2002 ◽  
Vol 322 (3) ◽  
pp. 543-557 ◽  
Author(s):  
Jane Greatorex ◽  
José Gallego ◽  
Gabriele Varani ◽  
Andrew Lever
Keyword(s):  
Wild Type ◽  
Packaging Signal ◽  
Structure And Stability ◽  
Internal Loops ◽  
Hiv 1
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