The cooperative function of arginine residues in the Prototype Foamy Virus Gag C-terminus mediates viral and cellular RNA encapsidation

Hamann, Martin V.; Müllers, Erik; Reh, Juliane; Stanke, Nicole; Effantin, Grégory; Weissenhorn, Winfríed; Lindemann, Dirk

doi:10.1186/preaccept-1342584341270573

Cited by 9 publications

(18 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Analysis of wt PFV ( Fig 6A and S2 Table ) reveals roughly spherical 1000 to 1300 Å diameter particles with external spikes of the Env protein and core structures as previously described [45, 54]. We performed cryo-tomography to study virus particles in 3-dimensions.…”

Section: Resultsmentioning

confidence: 84%

“…These seemingly incompatible data might be reconciled in the following way. It is known that initial FV capsid formation occurs within the cell cytoplasm and simultaneously viral RNA is recruited by Gag via the GR-regions [54]. Subsequently, FV Env leader peptide binds Gag to facilitate membrane targeting and particle release [45].…”

Section: Discussionmentioning

confidence: 99%

“…The particulate material was resuspended in phosphate-buffered saline (PBS). For cryo electron microscopy analysis viral particles were produced in serum-free medium and a further concentration step using Amicon Ultra 0.5 ml 100K Concentrators was included following the first concentration by ultracentrifugation through 20% sucrose similar as described recently [54]. …”

Section: Methodsmentioning

confidence: 99%

“…Transduction efficiency of recombinant, eGFP-expressing PFV vector particles by fluorescence marker-gene transfer assay was analyzed 72 h post-transduction as described previously [54, 95, 97]. All transduction experiments were performed at least twice.…”

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Structure of a Spumaretrovirus Gag Central Domain Reveals an Ancient Retroviral Capsid

et al. 2016

Self Cite

View full text Add to dashboard Cite

The Spumaretrovirinae, or foamy viruses (FVs) are complex retroviruses that infect many species of monkey and ape. Despite little sequence homology, FV and orthoretroviral Gag proteins perform equivalent functions, including genome packaging, virion assembly, trafficking and membrane targeting. However, there is a paucity of structural information for FVs and it is unclear how disparate FV and orthoretroviral Gag molecules share the same function. To probe the functional overlap of FV and orthoretroviral Gag we have determined the structure of a central region of Gag from the Prototype FV (PFV). The structure comprises two all α-helical domains NtDCEN and CtDCEN that although they have no sequence similarity, we show they share the same core fold as the N- (NtDCA) and C-terminal domains (CtDCA) of archetypal orthoretroviral capsid protein (CA). Moreover, structural comparisons with orthoretroviral CA align PFV NtDCEN and CtDCEN with NtDCA and CtDCA respectively. Further in vitro and functional virological assays reveal that residues making inter-domain NtDCEN—CtDCEN interactions are required for PFV capsid assembly and that intact capsid is required for PFV reverse transcription. These data provide the first information that relates the Gag proteins of Spuma and Orthoretrovirinae and suggests a common ancestor for both lineages containing an ancient CA fold.

show abstract

Section: Resultsmentioning

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Structure of a Spumaretrovirus Gag Central Domain Reveals an Ancient Retroviral Capsid

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…To assess the importance of GAG tethering to chromatin during infection, we produced singlecycle virus particles using codon-optimized constructs encoding PFV GAG (WT or R540Q), POL (WT or catalytically inert integrase mutant D185N/E221Q), and ENV along with a GFP reporter transfer vector (29). The vector particles were purified by ultracentrifugation through 20% sucrose cushions and quantified by immunoblotting using polyclonal anti-GAG antisera.…”

Section: Loss Of Cbs Function Leads To Catastrophic Redistribution Ofmentioning

confidence: 99%

Structural basis for spumavirus GAG tethering to chromatin

Lesbats

Serrao

Maskell

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

The interactions between a retrovirus and host cell chromatin that underlie integration and provirus expression are poorly understood. The prototype foamy virus (PFV) structural protein GAG associates with chromosomes via a chromatin-binding sequence (CBS) located within its C-terminal region. Here, we show that the PFV CBS is essential and sufficient for a direct interaction with nucleosomes and present a crystal structure of the CBS bound to a mononucleosome. The CBS interacts with the histone octamer, engaging the H2A-H2B acidic patch in a manner similar to other acidic patch-binding proteins such as herpesvirus latency-associated nuclear antigen (LANA). Substitutions of the invariant arginine anchor residue in GAG result in global redistribution of PFV and macaque simian foamy virus (SFV mac ) integration sites toward centromeres, dampening the resulting proviral expression without affecting the overall efficiency of integration. Our findings underscore the importance of retroviral structural proteins for integration site selection and the avoidance of genomic junkyards.retrovirus | integration | nucleosome | chromatin-binding

show abstract

TraFo-CRISPR: Enhanced Genome Engineering by Transient Foamy Virus Vector-Mediated Delivery of CRISPR/Cas9 Components

Lindel

Dodt

Weidner

et al. 2019

Molecular Therapy - Nucleic Acids

Self Cite

View full text Add to dashboard Cite

The adaptation of CRISPR/Cas technology for use in mammals has revolutionized genome engineering. In particular with regard to clinical application, efficient expression of Cas9 within a narrow time frame is highly desirable to minimize the accumulation of off-target editing. We developed an effective, aptamer-independent retroviral delivery system for Cas9 mRNAs that takes advantage of a unique foamy virus (FV) capability: the efficient encapsidation and transfer of non-viral RNAs. This enabled us to create a FV vector toolbox for efficient, transient delivery (TraFo) of CRISPR/Cas9 components into different target tissues. Co-delivery of Cas9 mRNA by TraFo-Cas9 vectors in combination with retroviral, integration-deficient single guide RNA (sgRNA) expression enhanced efficacy and specificity of gene-inactivation compared with CRISPR/Cas9 lentiviral vector systems. Furthermore, separate TraFo-Cas9 delivery allowed the optional inclusion of a repair matrix for efficient gene correction or tagging as well as the addition of fluorescent negative selection markers for easy identification of off-target editing or incorrect repair events. Thus, the TraFo CRISPR toolbox represents an interesting alternative technology for gene inactivation and gene editing.

show abstract

The cooperative function of arginine residues in the Prototype Foamy Virus Gag C-terminus mediates viral and cellular RNA encapsidation

Cited by 9 publications

References 47 publications

Structure of a Spumaretrovirus Gag Central Domain Reveals an Ancient Retroviral Capsid

Structure of a Spumaretrovirus Gag Central Domain Reveals an Ancient Retroviral Capsid

Structural basis for spumavirus GAG tethering to chromatin

TraFo-CRISPR: Enhanced Genome Engineering by Transient Foamy Virus Vector-Mediated Delivery of CRISPR/Cas9 Components

Contact Info

Product

Resources

About